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ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
additional information
?
-
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
480 ns multicopy molecular dynamics simulations performed, dynamics of apo and holo forms of the protein analyzed, opening and closing motions more pronounced in the apo form
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
crystal structure of BtuCD-F complex determined, BtuF protein shown to be bound to the periplasmic face of BtuCD
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
elastic normal mode analysis of BtuCD performed, mechanism of vitamin B12 transport cycle proposed
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
molecular dynamics on the vitamin B12-bound BtuF protein, energetics and mechanism of BtuF protein analyzed, opening and closing motions shown to be more pronounced in the apo form
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
nucleotide binding and release in the vitamin B12 importer BtuCD analyzed, perturbed elastic network calculations and biased molecular dynamics simulations applied, peristaltic forces suggested to exclude vitamin B12 from the transporter pore
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
simulation studies performed, analysis of principal components shown, with and without bound ATP
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
BtuCD is a type II ABC importer that catalyzes the translocation of vitamin B12 from the periplasm into the cytoplasm of Escherichia coli. BtuD is complexed with BtuC, a permease protein, and BtuF, a periplasmic binding protein, structure, overview
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-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
additional information
?
-
-
BtuCD alone or the BtuCD-F complex do not bind vitamin B12. Only free, uncomplexed BtuF binds vitamin B12 with high affinity. Upon binding of BtuF to BtuCD, vitamin B12 is released from BtuF and is only transiently associated with the complex
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-
?
additional information
?
-
-
BtuCD forms a stable complex with the vitamin B12 binding protein BtuF. Using protein docking and MD simulation studies it is shown that holo-BtuF stabilizes the open conformation of BtuCD, whereas the transporter begins to close again when BtuF or vitamin B12 is removed suggesting BtuCD-F is capable of substrate sensitivity. BtuC transmembrane helices 3 and 5, the L-loops and the adjacent helices comprised of BtuC residues 170-180 are identified as hotspots of conformational change
-
-
?
additional information
?
-
-
BtuCD forms an extremely stable complex with the vitamin B12 binding protein BtuF. Vitamin B12 accelerates complex dissociation rate, with ATP having an additional destabilizing effecf
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-
?
additional information
?
-
-
in the presence of vitamin B12, upon binding of ATP (or ATP analogues), no association between BtuF and BtuCD can be detected
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-
?
additional information
?
-
a vitamin B12 molecule remains bound to the liposome-reconstituted transporter complex for tens of seconds, during which several ATP hydrolysis cycles can take place, before it is being transported across the membrane. Measurements of fluorescence changes on BtuCD induced by BtuF and ATP, transport of single vitamin B12 molecules, overview
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-
?
additional information
?
-
BtuCD-F catalyzes the uptake of cobinamide, a cobalamin precursor, and cobalamin. BtuCD-catalyzed in vitro transport of cyano-cobinamide and of cobalamin is ATP- and BtuF-dependent. Tryptophan residue W66 of BtuF is involved in the substrate binding of cobalamin
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?
additional information
?
-
-
BtuCD-F catalyzes the uptake of cobinamide, a cobalamin precursor, and cobalamin. BtuCD-catalyzed in vitro transport of cyano-cobinamide and of cobalamin is ATP- and BtuF-dependent. Tryptophan residue W66 of BtuF is involved in the substrate binding of cobalamin
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-
?
additional information
?
-
-
metal-chelate-type ABC transporter HI1470/1 is homologous with vitamin B12 importer BtuCD but exhibits a distinct inward-facing conformation in contrast to the outward-facing conformation of BtuCD. The outward-facing conformation of HI1470/1 is considered to be one of the intermediate states in the translocation cycle of BtuCD
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?
additional information
?
-
the enzyme possesses equal levels of acyl-CoA thioesterase activity, ATPase and thioesterase activities of liposome-reconstituted human ABCD4, overview
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-
?
additional information
?
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-
the enzyme possesses equal levels of acyl-CoA thioesterase activity, ATPase and thioesterase activities of liposome-reconstituted human ABCD4, overview
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-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
hemin does not compete with cobalamin-uptake. Although promiscuous among cobalamin variants and cobalamin precursors, ECF-CbrT is a dedicated vitamin B12 transporter
-
-
?
additional information
?
-
BtuM binds vitamin B12 in its base-off conformation, in which the 5,6-dimethylbenzimidazole moiety does not bind to the cobalt ion, but with a cysteine residue as axial ligand of the corrin cobalt ion, BtuMTd binds cobalamin (Cbl) using cysteine ligation. In contrast, at physiological pH the conformation of free Cbl in aqueous solution is base-on with the 5,6-dimethylbenzimidazole moiety coordinated to the cobalt ion in the alpha-axial position
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?
additional information
?
-
BtuM binds vitamin B12 in its base-off conformation, in which the 5,6-dimethylbenzimidazole moiety does not bind to the cobalt ion, but with a cysteine residue as axial ligand of the corrin cobalt ion, BtuMTd binds cobalamin (Cbl) using cysteine ligation. In contrast, at physiological pH the conformation of free Cbl in aqueous solution is base-on with the 5,6-dimethylbenzimidazole moiety coordinated to the cobalt ion in the alpha-axial position. The membrane environment also appears to preclude Cbl binding to purified BtuMTd, as binding is observed only when the substrate is added before solubilisation. BtuMTd also catalyses cysteine-mediated decyanation of vitamin B12
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-
?
additional information
?
-
VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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-
?
additional information
?
-
VcBtuF shows a distinctive phenomenon of heme binding with comparable affinity to vitamin B12
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-
?
additional information
?
-
VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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-
?
additional information
?
-
VcBtuF shows a distinctive phenomenon of heme binding with comparable affinity to vitamin B12
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-
?
additional information
?
-
VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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-
?
additional information
?
-
VcBtuF shows a distinctive phenomenon of heme binding with comparable affinity to vitamin B12
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-
?
additional information
?
-
VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
-
-
?
additional information
?
-
VcBtuF shows a distinctive phenomenon of heme binding with comparable affinity to vitamin B12
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
additional information
?
-
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + cobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + cyanocobalamin-[cobalamin-binding protein][side 1]
ADP + phosphate + cyanocobalamin[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + dicyanocobinamide-[cobalamin-binding protein][side 1]
ADP + phosphate + dicyanocobinamide[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12-[cobalamin-binding protein][side 1]
ADP + phosphate + vitamin B12[side 2] + [cobalamin-binding protein][side 1]
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
BtuCD is a type II ABC importer that catalyzes the translocation of vitamin B12 from the periplasm into the cytoplasm of Escherichia coli. BtuD is complexed with BtuC, a permease protein, and BtuF, a periplasmic binding protein, structure, overview
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
ATP + H2O + vitamin B12/out
ADP + phosphate + vitamin B12/in
-
-
-
-
?
additional information
?
-
-
BtuCD alone or the BtuCD-F complex do not bind vitamin B12. Only free, uncomplexed BtuF binds vitamin B12 with high affinity. Upon binding of BtuF to BtuCD, vitamin B12 is released from BtuF and is only transiently associated with the complex
-
-
?
additional information
?
-
-
BtuCD forms a stable complex with the vitamin B12 binding protein BtuF. Using protein docking and MD simulation studies it is shown that holo-BtuF stabilizes the open conformation of BtuCD, whereas the transporter begins to close again when BtuF or vitamin B12 is removed suggesting BtuCD-F is capable of substrate sensitivity. BtuC transmembrane helices 3 and 5, the L-loops and the adjacent helices comprised of BtuC residues 170-180 are identified as hotspots of conformational change
-
-
?
additional information
?
-
-
BtuCD forms an extremely stable complex with the vitamin B12 binding protein BtuF. Vitamin B12 accelerates complex dissociation rate, with ATP having an additional destabilizing effecf
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additional information
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in the presence of vitamin B12, upon binding of ATP (or ATP analogues), no association between BtuF and BtuCD can be detected
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additional information
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metal-chelate-type ABC transporter HI1470/1 is homologous with vitamin B12 importer BtuCD but exhibits a distinct inward-facing conformation in contrast to the outward-facing conformation of BtuCD. The outward-facing conformation of HI1470/1 is considered to be one of the intermediate states in the translocation cycle of BtuCD
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additional information
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BtuM binds vitamin B12 in its base-off conformation, in which the 5,6-dimethylbenzimidazole moiety does not bind to the cobalt ion, but with a cysteine residue as axial ligand of the corrin cobalt ion, BtuMTd binds cobalamin (Cbl) using cysteine ligation. In contrast, at physiological pH the conformation of free Cbl in aqueous solution is base-on with the 5,6-dimethylbenzimidazole moiety coordinated to the cobalt ion in the alpha-axial position
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additional information
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VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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additional information
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VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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additional information
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VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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additional information
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VcBtuF, the periplasmic substrate binding protein (PBP) of putative ABC transporter BtuF-CD of Vibrio cholerae O395, binds cyanocobalamin and dicyanocobinamide with micromolar dissociation constants (Kd). Productive internalization of these nutrients
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0.18
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in the absence of Btu-F and vitamin B12 in proteoliposomes
0.33
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in the presence of Fos-choline
0.44
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in the presence of 0.02 mmol BtuF in proteoliposomes
0.98
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in the presence of lauryl dimethylamine-N-oxide
additional information
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vitamin B12 has little effect on the activity in proteoliposomes, as well as in the presence of lauryl dimethylamine-N-oxide, dodecyl maltoside and Triton X-100
additional information
crystal structure of BtuCD-F complex presented, displacement of vitamin B12 from binding pocket shown, subunits of transmembrane BtuC protein shown to have two distinct conformations, translocation pathway shown to be closed to both sides of the membrane, electron paramagnetic resonance spectra of spin-labeled cysteine mutants shown to be consistent with the conformation of BtuCD-F observed in the crystal structure, structure of BtuCD-F discussed as a posttranslocation intermediate
additional information
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crystal structure of BtuCD-F complex presented, displacement of vitamin B12 from binding pocket shown, subunits of transmembrane BtuC protein shown to have two distinct conformations, translocation pathway shown to be closed to both sides of the membrane, electron paramagnetic resonance spectra of spin-labeled cysteine mutants shown to be consistent with the conformation of BtuCD-F observed in the crystal structure, structure of BtuCD-F discussed as a posttranslocation intermediate
additional information
intrinsic flexibility of isolated BtuC and BtuD dimers evaluated, conformational movement of intact BtuCD transporter studied in the context of conformational coupling between BtuC and BtuD, effect of BtuF association on the transporter conformational movement discussed
additional information
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intrinsic flexibility of isolated BtuC and BtuD dimers evaluated, conformational movement of intact BtuCD transporter studied in the context of conformational coupling between BtuC and BtuD, effect of BtuF association on the transporter conformational movement discussed
additional information
perturbed anisotropic network model and molecular dynamics simulations applied, essential dynamics sampling and cavity and pathway analysis, structural response of BtuC protein quantified in terms of X-shifts shown, transport model discussed
additional information
principal motions and conformational changes of apo and holo forms studied, closed conformation for the ligand-bound state identified, empty form shown to fluctuate between open and closed conformations, simulation system and force field parameters described, elastic network normal-mode analysis performed, conformational changes summarized, domain motions indicated, link to previous simulations and biological implications discussed
additional information
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principal motions and conformational changes of apo and holo forms studied, closed conformation for the ligand-bound state identified, empty form shown to fluctuate between open and closed conformations, simulation system and force field parameters described, elastic network normal-mode analysis performed, conformational changes summarized, domain motions indicated, link to previous simulations and biological implications discussed
additional information
principal motions of BtuCD-F transporter system and conformation of ATP-binding sites analyzed, protein subunits representing simulation components indicated, design of simulation system described, conformational drift summarized, model of the BtuCD-F complex discussed
additional information
steered molecular dynamics simulations performed, principal component analysis and energetics of vitamin B12 unbinding analyzed, potential of mean force along postulated vitamin B12 unbinding pathway constructed through Jarzynski's equality, motion modes and intrinsic flexibility of BtuF calculated, Trp44-Gln45 pair situated at binding pocket suggested to act as a gate in the vitamin B12 binding and unbinding process
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evolution
ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
evolution
Escherichia coli vitamin B12 transporter BtuCD-F is a type II importer and belongs to the ATP-binding cassette (ABC) transporter superfamily
evolution
the enzyme is a ABC transporter, all ABC-type ATPases encoded by the organism are predicted to be part of classical ABC transporters, and not ECF transporters. The organism does not encode an ECF-module. BtuM homologues (apart from one exception) are found exclusively in organisms lacking an ECF-module. But BtuMTd structurally resembles the S-components of ECF transporters
evolution
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ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
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evolution
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ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
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evolution
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ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
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evolution
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ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
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evolution
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ECF-transporters are multi-subunit membrane complexes that consist of two ATPases, similar to the ATPases of ABC transporters, and two membrane embedded proteins, not related to any other protein family
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malfunction
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gene disruption eliminates the ability of Mycobacterium tuberculosis to use exogenous vitamin B12 in vitro
malfunction
ABCD4 dysfunction results in a failure of lysosomal vitamin B12 release
malfunction
mutations in ABCD4 result in a failure to release vitamin B12 from lysosomes. A similar phenotype is caused by mutations in gene LMBRD1, which encodes the lysosomal membrane protein LMBD1. ABCD4 lysosomal localization is disturbed by knockout of LMBRD. Mutation of ABCD4, which is known as the cblJ complementation group, results in the failure of the release of cobalamin from lysosomes. A similar phenotype in patients within the cblF group is caused by mutations of LMBD1, a lysosomal membrane protein. Mistargeting of mutant LMBD1 affects the distribution of ABCD4. Thus, mutations of ABCD4 and LMBD1 result in a quite similar phenotype. A putative region of ABCD4 that interacts with LMBD1 might be masked by the exchange of the regions around transmembrane domains (TMs) 2 and 5
malfunction
substitution of W66 in BtuF with tyrosine or leucine reduced the affinity 3fold compared to wild-type, and a change to histidine or arginine reduces it more than 10fold
physiological function
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the enzyme is required for the assimilation of exogenous vitamin B12 to enable methylmalonyl-CoA pathway function and is essential for corrinoid transport
physiological function
ABCD4 is a transporter of cobalamin and forms a complex with LMBD1 for the proper targeting or functioning, or both. The two proteins function as a complex
physiological function
ABCD4 is located on lysosomal membrane and is involved in the transport of vitamin B12 from lysosomes to the cytosol
physiological function
ATP-binding cassette (ABC) transporters are a large family of integral membrane proteins and involved in nutrient uptake, drug extrusion, and lipid homeostasis. They use the energy of ATP binding and hydrolysis to power substrate transport across the lipid bilayer. BtuCD-F is an ABC transporter that mediates cobalamin (Cbl) uptake into Escherichia coli, Escherichia coli is unable to synthesize Cbl de novo. BtuCD-F might also be involved in the uptake of cobinamide, a cobalamin precursor. Precursor cobinamide (Cbi) lacks the 5,6-dimethylbenzimidazole (DMB) moiety and sugar-phosphate linker and is therefore smaller than Cbl. BtuCD-catalyzed in vitro transport of cyano-cobinamide is ATP- and BtuF-dependent. BtuF residue W66 is important for high affinity Cbi binding, but not for substrate delivery or transport
physiological function
ATP-binding cassette (ABC) transporters form the largest class of active membrane transport proteins. Binding and hydrolysis of ATP by their highly conserved nucleotide-binding domains drive conformational changes of the complex that mediate transport of substrate across the membrane. The transporter complex of vitamin B12 importer BtuCD-F from Escherichia coli is consisting of a periplasmic soluble binding protein BtuF that binds the ligand and the transmembrane and ATPase domains BtuCD mediating translocation
physiological function
bacterial ABC importers catalyze the uptake of essential nutrients including transition metals and metal-containing cofactors
physiological function
cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
physiological function
the ABC transporter BtuCD-F imports vitamin B12 across the inner membrane of Escherichia coli. Substrate translocation by ATP-binding cassette (ABC) transporters involves coupling of ATP binding and hydrolysis in the nucleotide-binding domains (NBDs) to conformational changes in the transmembrane domains
physiological function
the ATP-binding cassette (ABC) importer family catalyzse the uptake of nutrients, vitamins and trace elements. Membrane transport proteins generally are inherently flexible and undergo substantial conformational changes to catalyze the translocation of their substrates across biological membranes. The vitamin B12 import system BtuCD-F from Escherichia coli shows the conformational dynamics during the transport cycle
physiological function
the membrane protein BtuM acts as transporter for uptake of essential vitamin B12, i.e. cobalamin, one of the most complex cofactors known, and used by enzymes catalyzing for instance methyl-group transfer and ribonucleotide reduction reactions. BtuMTd likely combines two functions: transport of the substrate into the bacterial cell, and chemical modification of the substrate. A cobalt-cysteine interaction allows for chemical modification of the substrate prior to translocation, which is a rare feature among uptake systems. BtuM homologues are small membrane proteins of about 22 kDa, and found predominantly in Gram-negative species, distributed mostly over alpha-, beta-, and gamma-proteobacteria
physiological function
vitamin B12 importer BtuCD is a type II ATP binding cassette (ABC) importer mediating the uptake of vitamin B12 across the inner membrane. ABC transporters utilize the energy of ATP hydrolysis to unidirectionally transport substrates across cell membrane. ATP hydrolysis occurs at the nucleotide-binding domain (NBD) dimer interface of ABC transporters, whereas substrate translocation takes place at the translocation pathway between the transmembrane domains (TMDs), which is more than 30 A away from the NBD dimer interface
physiological function
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cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
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physiological function
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cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
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physiological function
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cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
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physiological function
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cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
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physiological function
-
cobalamin-specific ECF-type ABC transporter from Lactobacillus delbrueckii, ECF-CbrT, mediates the specific, ATP-dependent uptake of cobalamin. Cobalamin (vitamin B12) is the most complex B-type vitamin and is synthetized exclusively in a limited number of prokaryotes. Its biologically active variants contain rare organometallic bonds, which are used by enzymes in a variety of central metabolic pathways such as L-methionine synthesis and ribonucleotide reduction. Enzyme ECF-CbrT catalyzes ATP-dependent transport of cobalamin and cobinamide
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additional information
ABC importers follow the two-site access model3, in which ATP binding and hydrolysis switch the accessibility of the transmembrane domain for the substrate from an inward facing (accessible from the cytoplasm) to an outward-facing (accessible from the extracellular site) conformation, conformational changes by single-molecule FRET measurements combined with molecular dynamics simulations, two different transport cycles are analyzed
additional information
conformational coupling, molecular dynamics simulations using BtuCD-F is embedded in a solvated phosphatidylcholine bilayer, configurational entropy, pairwise residue-residue forces in BtuCD-F are analyzed through force distribution analysis, overview
additional information
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conformational coupling, molecular dynamics simulations using BtuCD-F is embedded in a solvated phosphatidylcholine bilayer, configurational entropy, pairwise residue-residue forces in BtuCD-F are analyzed through force distribution analysis, overview
additional information
function and structure of BtuMTd, cobalamin binding structure, overview
additional information
molecular dynamics simulation of structure of the cobalamin-binding protein BtuF compared to Escherichia coli BtuF structure
additional information
post-hydrolysis state of the vitamin B12 importer BtuCD by molecular dynamics (MD) simulations, overview. Predominantly asymmetric arrangement of the NBD dimer interface, with the ADP-bound site disrupted and the ATP-bound site preserved in most of the trajectories. TMDs response to ATP hydrolysis by separation of the L-loops and opening of the cytoplasmic gate II, indicating that hydrolysis of one ATP facilitates substrate translocation by opening the cytoplasmic end of translocation pathway. Motions of the L-loops and the cytoplasmic gate II are coupled with each other through a contiguous interaction network involving a conserved Asn83 on the extended stretch preceding transmembrane (TM)3 helix plus the cytoplasmic end of TM2/6/7 helix bundle. TMD-NBD communication mechanism for type II ABC importers. Besides the four basic domains of BtuCD, a cognate periplasmic binding protein, BtuF, is also required to maximize transport rate. Different conformational states of BtuCD, and mechanism of B12 transport cycle in BtuCD, overview. The occluded state of BtuCD, occ-BtuCD (PDB ID 4FI3), is regarded as a crucial step of the transport cycle, in which the transporter simultaneously loads the shipment B12 and the energy source ATPs. Transition from the occ-BtuCD state to the inward-facing state after ATP hydrolysis
additional information
the crystal structure of cobinamide-bound BtuF reveals a conformational change of a tryptophan residue W66 in the substrate binding cleft compared to the structure of cobalamin-bound BtuF, molecular dynamics simulations. BtuF is a class III periplasmic substrate binding protein
additional information
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the crystal structure of cobinamide-bound BtuF reveals a conformational change of a tryptophan residue W66 in the substrate binding cleft compared to the structure of cobalamin-bound BtuF, molecular dynamics simulations. BtuF is a class III periplasmic substrate binding protein
additional information
the homodimer BtuC spans the membrane and the two identical cytosolic ATPase domains BtuD form a sandwich dimer that couple chemical energy of two ATP molecules into structural changes of the full complex. A single substrate-binding protein (SBP) completes the transporter. The SBP belongs to cluster A or class III and exhibits relatively small conformational changes upon substrate binding. Modeling of the transport mechanism of BtuCD-F transporters embedded in lipid bilayers at the single molecule level, overview
additional information
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molecular dynamics simulation of structure of the cobalamin-binding protein BtuF compared to Escherichia coli BtuF structure
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additional information
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molecular dynamics simulation of structure of the cobalamin-binding protein BtuF compared to Escherichia coli BtuF structure
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additional information
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molecular dynamics simulation of structure of the cobalamin-binding protein BtuF compared to Escherichia coli BtuF structure
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analysis of the cobinamide (Cbi)-bound BtuF crystal structure model, PDB ID 5M29, crystal structures of Cbi-bound BtuF mutants W66F, W66Y and W66L, sitting drop vapor diffusion technique, mixing of 20 mg/ml protein in 10 mM Tris pH 8 and 100 mM NaCl, with precipitant solution containing 1% w/v tryptone, 50 mM HEPES, pH 7.0, and 12% w/v PEG 3350, 1-2 weeks, 20°C, X-ray diffraction structure determination and analysis at 1.5-1.7 A resolution, molecular replacement using the BtuF structure (PDB ID 1N2Z) as search model
BtuCD-F complex analyzed at a resolution of 2.6 A, substantial conformational changes observed as compared with previously reported structures of BtuCD and BtuF
catalytically impaired BtuCD mutant E159Q in complex with BtuF, to 3.5 A resolution. The BtuC subunits adopts a distinct asymmetric conformation. The structure suggests that BtuF does not discriminate between, or impose, asymmetric conformations of BtuCD
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crystal structure analysis of the crystal structure of the BtuCD-F complex, modelling, overview
elucidation of gating mechanism by EPR spectroscopy. The translocation gates of the BtuCDF complex undergo conformational changes in line with a two-state alternating access model. Binding of ATP drives the gates to an inward-facing conformation. Following ATP hydrolysis, the translocation gates restore to an apo-like conformation. In the presence of ATP, an excess of vitamin B12 promotes the reopening of the gates toward the periplasm and the dislodgement of BtuF from the transporter
in complex with beta-gamma-imidoadenosine 5'-triphosphate, sitting drop vapor diffusion method, using 20-30% (w/v) PEG 400, 100 mM N-(2-acetamido)-iminodiacetic acid, pH 6.8, 100 mM sodium potassium citrate
molecular dynamics simulations to explore the atomic details of the conformational transitions of BtuCD importer. The outward-facing to inward-facing transition is initiated by the conformational movement of nucleotide-binding domains. The subsequent reorientation of the substrate translocation pathway at transmembrane domains begins with the closing of the periplasmic gate, followed by the opening of the cytoplasmic gate in the last stage of the conformational transition due to the extensive hydrophobic interactions at this region, consistent with the functional requirement of unidirectional transport of the substrates. The reverse inward-facing to outward-facing transition exhibits intrinsic diversity of the conformational transition pathways and significant structural asymmetry
mutant E159Q/N162C in complex with adenylyl imidodiphosphate, sitting drop vapor diffusion method, using 100 mM ADA buffer, pH 6.9, 1.2 M NaCl, and 14-18% (w/v) PEG 2000 MME
purified recombinant complex of inhibitory nanobody Nb9 with enzyme BtuF complex, crystallization solution contains 100 mM Tris-HCl, pH 8.5, 400 mM MgCl2, and 33% w/v PEG4000, X-ray diffraction structrue determination and analysis at 2.7 A resolution
purified recombinant wild-type BtuCD-F, apo-BtuCD-F, and BtuCD-F mutant E159Q/N162C, X-ray diffraction structure determination and analysis
BtuCD-F complex, and HI470/1 X-ray diffraction structure analysis using PDB-IDs 2NQ2, 1L7V and 2QI9, overview
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purified ECF-CbrT complex in detergent (n-dodecyl-beta-D-maltopyranoside) solution, sitting drop vapour diffusion method, mixing of protein solution containing 50 mM HEPES pH 8, 150 mM NaCl, 1% polyoxyethylene(10)dodecyl ether, with precipitant solution containing 0.2 M KCl, 0.1 M sodium citrate, pH 5.5, 37% v/v pentaerythritol propoxylate, X-ray diffraction structure determination and analysis at 3.4 A resolution, molecular replacement with the structure of the folate transporter, ECF-FolT2, from Lactobacillus delbrueckii as a search model
purified recombinant His-tagged enzyme BtuM with natively bound cobalmin and anomalously bound cobalmin, sitting drop vapour diffusion method, mixing of 0.002 ml of protein in 50mM Tris-HCl, pH 7.5, or in 50 mM HEPES-NaOH pH 8.0, 100 mM NaCl, 0.005 mM cyano-Cbl and 0.35% detergent, with 0.002 ml of precipitant solution containing 25 mM Tris, pH 8.5, and 25-30% v/v PEG 400 or 50 mM Tris, pH 8.5, and 27-30% v/v PEG 400, or 75 mM Tris, pH 8.5, and 29-30% v/v PEG 400, at 8°C, 3-4 weeks, X-ray diffraction structure determination and analysis at 2.0-2.5 A resolution, structure modeling
vitamin B12-bound VcBtuF, protein in a solution with vitamin B12 in a 3:1 ratio, and 50 mM Tris-HCl, pH 7.0, and 300 mM NaCl, mixing of 0.003 ml of protein solution with 0.002 ml of precipitant solution containing 0.8 M ammonium sulfate, 0.1 M Tris, pH 8.0, and equilibration against a reservoir solution containing 0.5 ml of 1.6 M ammonium sulfate, 0.1 M HEPES, pH 7.0, 20°C, 7 days, hanging drop vapour diffusion method, X-ray diffraction structure determination and analysis at 1.67 A resolution
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D141C
site-directed mutagenesis of BtuF on a residue pointing outward in the middle of the alpha-helix connecting the two lobes, the mutation allows for specific coupling of fluorescent labels to each of these proteins
L85C
the mutant shows a 6.5fold reduction in ATPase activity compared to the wild type enzyme
Q111C
site-directed mutagenesis of BtuC on the periplasmic loop connecting transmembrane (TM) helix 3 and 4
S141C
site-directed mutagenesis, structure analysis
S143C
site-directed mutagenesis, structure analysis
T168C
site-directed mutagenesis, structure analysis
W115L
site-directed mutagenesis of BtuC to remove any quenching effects of this tryptophan on the fluorescent probes
W66A
site-directed mutagenesis, reduces the affinity for cobinamide severalfold compared to wild-type
W66E
site-directed mutagenesis, reduces the affinity for cobinamide severalfold compared to wild-type
W66F
site-directed mutagenesis, does not reduce the affinity for cobinamide severalfold compared to wild-type
W66H
site-directed mutagenesis, reduces the affinity for cobinamide 10fold compared to wild-type
W66L
site-directed mutagenesis, reduces the affinity for cobinamide 3fold compared to wild-type
W66R
site-directed mutagenesis, reduces the affinity for cobinamide 10fold compared to wild-type
W66Y
site-directed mutagenesis, reduces the affinity for cobinamide severalfold compared to wild-type
E159Q
-
mutation in subunit BtuD, results in abolished ATP hydrolysis activity of BtuCDF. Mutant is still able to bind nucleotides and binding protein BtuF in a manner similar to the wild-type protein
E159Q
site-directed mutagenesis of BtuD, an ATPase impaired mutant
E159Q/N162C
the mutant is unable to transport substrate despite a very low residual ATP hydrolysis rate
E159Q/N162C
the mutant shows dramatically reduced ATPase activity
E159Q/N162C
site-directed mutagenesis, a disulfide mutant, analysis of the crystal structure of the mutant with bound ATP
additional information
-
a system consisting of the BtuC subunit embedded in a palmitoyloleoyl phosphatidylcholine lipid bilayer is constructed and a more-than-57ns MD simulation is performed to study the functional motions of BtuC at the atomic level: results show that a stable protein-lipid bilayer is obtained and the palmitoyloleoyl phosphatidylcholine lipid bilayer is able to adjust its thickness to match the embedded BtuC which undergo relatively complicated motions
additional information
site-directed mutagenesis of tryptophan residue W66 in the substrate binding cleft , the affinity for cobinamide of the W66X mutants is lower except for W66F. Three mutants with impaired Cbi binding (W66A, W66R, and W66E) and one with high binding affinity (W66F) are used for transport assays. Despite having lower Cbi binding affinities, Cbi transport is hardly affected by W66X substitution
additional information
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site-directed mutagenesis of tryptophan residue W66 in the substrate binding cleft , the affinity for cobinamide of the W66X mutants is lower except for W66F. Three mutants with impaired Cbi binding (W66A, W66R, and W66E) and one with high binding affinity (W66F) are used for transport assays. Despite having lower Cbi binding affinities, Cbi transport is hardly affected by W66X substitution
additional information
the mutations and labels on BtuF and BtuC have no critical effect on ATP hydrolysis and transport activity
additional information
generation of chimeric ABCD4 proteins that are exchanged in terms of the corresponding putative transmembrane helix with ABCD1 (based on the secondary structure of the eukaryotic P-glycoprotein homolog CmABCB1) in HEK-293 cells, endogenous human ABCD1 does not interact with LMBD1. Construction of ABCD4 chimeras 1-6 and analysis of the localization of chimeric ABCD4s in CHO cells stably expressing LMBD1-GFP. The wild-type ABCD4 co-expressed with LMBD1 exhibits a punctate distribution that is superimposable on the distribution pattern of LMBD1. The distribution patterns of the ABCD4 chimeras 1, 3, 4 and 6 also display the same pattern as LMBD1. But ABCD4 chimeras 2 and 5 do not exhibit a punctate pattern, but rather, a reticulum-like distribution pattern that is not superimposable on LMBD1
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Friedrich, M.J.; DeVaux, L.C.; Kadner,R.
Nucleotide sequence of the btuCED genes involved in vitamin B12 transport in Escherichia coli and homology with components of periplasmic-binding-protein-dependent transport systems
J. Bacteriol.
167
928-934
1986
Escherichia coli
brenda
Kadner, R.J.
Vitamin B12 transport in Escherichia coli: energy coupling between membranes
Mol. Microbiol.
12
2027-2033
1990
Escherichia coli
brenda
Rioux, C.R.; Kadner, R.J.
Vitamin B12 transport in Escherichia coli K12 does not require the btuE gene of the Btu CED operon
Mol. Gen. Genet.
217
301-308
1989
Escherichia coli
brenda
De Vaux, L.C.; Clevenson, D.S.; Bradbeer, C.; Kadner, R.J.
Identification of the btuCED polypeptides and evidence for their role in vitamin B12 transport in Escherichia coli
J. Bacteriol.
167
920-927
1986
Escherichia coli
brenda
De Vaux, L.C.; Kadner, R.
Transport of vitamin B12 in Escherichia coli: Cloning of the btuCD region
J. Bacteriol.
162
888-896
1985
Escherichia coli
brenda
Rioux, C.R.; Kadner, R.J.
Two outer membrane transport systems for vitamin B12 in Salmonella typhimurium
J. Bacteriol.
171
2986-2993
1989
Salmonella enterica subsp. enterica serovar Typhimurium
brenda
Borths, E.L.; Poolman, B.; Hvorup, R.N.; Locher, K.P.; Rees, D.C.
In vitro functional characterization of BtuCD-F, the Escherichia coli ABC transporter for vitamin B12 uptake
Biochemistry
44
16301-16309
2005
Escherichia coli
brenda
Kandt, C.; Xu, Z.; Tieleman, D.P.
Opening and closing motions in the periplasmic vitamin B12 binding protein BtuF
Biochemistry
45
13284-13292
2006
Escherichia coli (P06611), Escherichia coli
brenda
Ivetac, A.; Campbell, J.D.; Sansom, M.S.
Dynamics and function in a bacterial ABC transporter: simulation studies of the BtuCDF system and its components
Biochemistry
46
2767-2778
2007
Escherichia coli (P06611)
brenda
Liu, M.; Sun, T.; Hu, J.; Chen, W.; Wang, C.
Study on the mechanism of the BtuF periplasmic-binding protein for vitamin B(12)
Biophys. Chem.
135
19-24
2008
Escherichia coli (P37028)
brenda
Sonne, J.; Kandt, C.; Peters, G.H.; Hansen, F.Y.; Jensen, M.?.; Tieleman, D.P.
Simulation of the coupling between nucleotide binding and transmembrane domains in the ATP binding cassette transporter BtuCD
Biophys. J.
92
2727-2734
2007
Escherichia coli (P37028)
brenda
Weng, J.; Ma, J.; Fan, K.; Wang, W.
The conformational coupling and translocation mechanism of vitamin B12 ATP-binding cassette transporter BtuCD
Biophys. J.
94
612-621
2008
Escherichia coli (P06611), Escherichia coli
brenda
Hvorup, R.N.; Goetz, B.A.; Niederer, M.; Hollenstein, K.; Perozo, E.; Locher, K.P.
Asymmetry in the structure of the ABC transporter-binding protein complex BtuCD-BtuF
Science
317
1387-1390
2007
Escherichia coli (P06611), Escherichia coli
brenda
Weng, J.; Ma, J.; Fan, K.; Wang, W.
Asymmetric conformational flexibility in the ATP-binding cassette transporter HI1470/1
Biophys. J.
96
1918-1930
2009
Haemophilus influenzae
brenda
Goetz, B.A.; Perozo, E.; Locher, K.P.
Distinct gate conformations of the ABC transporter BtuCD revealed by electron spin resonance spectroscopy and chemical cross-linking
FEBS Lett.
583
266-270
2009
Escherichia coli (P06611), Escherichia coli
brenda
Lewinson, O.; Lee, A.T.; Locher, K.P.; Rees, D.C.
A distinct mechanism for the ABC transporter BtuCD-BtuF revealed by the dynamics of complex formation
Nat. Struct. Mol. Biol.
17
332-338
2010
Escherichia coli
brenda
Kandt, C.; Tieleman, D.P.
Holo-BtuF stabilizes the open conformation of the vitamin B12 ABC transporter BtuCD
Proteins
78
738-753
2010
Escherichia coli
brenda
Sun, T.; Liu, M.; Chen, W.; Wang, C.
Molecular dynamics simulation of the transmembrane subunit of BtuCD in the lipid bilayer
Sci. China C Life Sci.
53
620-630
2010
Escherichia coli
brenda
Korkhov, V.M.; Mireku, S.A.; Hvorup, R.N.; Locher, K.P.
Asymmetric states of vitamin B12 transporter BtuCD are not discriminated by its cognate substrate binding protein BtuF
FEBS Lett.
586
972-976
2012
Escherichia coli
brenda
Di Bartolo, N.D.; Hvorup, R.N.; Locher, K.P.; Booth, P.J.
In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD
J. Biol. Chem.
286
18807-18815
2011
Escherichia coli
brenda
Joseph, B.; Jeschke, G.; Goetz, B.A.; Locher, K.P.; Bordignon, E.
Transmembrane gate movements in the type II ATP-binding cassette (ABC) importer BtuCD-F during nucleotide cycle
J. Biol. Chem.
286
41008-41017
2011
Escherichia coli (P06609), Escherichia coli
brenda
Weng, J.; Fan, K.; Wang, W.
The conformational transition pathways of ATP-binding cassette transporter BtuCD revealed by targeted molecular dynamics simulation
PLoS ONE
7
e305465
2012
Escherichia coli (P06611)
-
brenda
Su, J.G.; Zhang, X.; Zhao, S.X.; Li, X.Y.; Hou, Y.X.; Wu, Y.D.; Zhu, J.Z.; An, H.L.
Conformational motions and functionally key residues for vitamin B12 transporter BtuCD-BtuF revealed by elastic network model with a function-related internal coordinate
Int. J. Mol. Sci.
16
17933-17951
2015
Escherichia coli (P37028), Escherichia coli
brenda
Joseph, B.; Korkhov, V.M.; Yulikov, M.; Jeschke, G.; Bordignon, E.
Conformational cycle of the vitamin B12 ABC importer in liposomes detected by double electron-electron resonance (DEER)
J. Biol. Chem.
289
3176-3185
2014
Escherichia coli
brenda
Korkhov, V.M.; Mireku, S.A.; Veprintsev, D.B.; Locher, K.P.
Structure of AMP-PNP-bound BtuCD and mechanism of ATP-powered vitamin B12 transport by BtuCD-F
Nat. Struct. Mol. Biol.
21
1097-1099
2014
Escherichia coli (P06609), Escherichia coli
brenda
Korkhov, V.M.; Mireku, S.A.; Locher, K.P.
Structure of AMP-PNP-bound vitamin B12 transporter BtuCD-F
Nature
490
367-372
2012
Escherichia coli (P06609), Escherichia coli
brenda
Gopinath, K.; Venclovas, A.; Ioerger, T.; Sacchettini, J.; McKinney, J.; Mizrahi, V.; Warner, D.
A vitamin B12 transporter in Mycobacterium tuberculosis
Open Biology
3
120175
2013
Mycobacterium tuberculosis
brenda
Okamoto, T.; Kawaguchi, K.; Watanabe, S.; Agustina, R.; Ikejima, T.; Ikeda, K.; Nakano, M.; Morita, M.; Imanaka, T.
Characterization of human ATP-binding cassette protein subfamily D reconstituted into proteoliposomes
Biochem. Biophys. Res. Commun.
496
1122-1127
2018
Homo sapiens (O14678), Homo sapiens
brenda
Agarwal, S.; Dey, S.; Ghosh, B.; Biswas, M.; Dasgupta, J.
Mechanistic basis of vitamin B12 and cobinamide salvaging by the Vibrio species
Biochim. Biophys. Acta
1867
140-151
2019
Vibrio cholerae serotype O1 (A0A0H3AMA6 AND A5F1V0 AND A5F5P5), Vibrio cholerae serotype O1 Classical Ogawa 395 (A0A0H3AMA6 AND A5F1V0 AND A5F5P5), Vibrio cholerae serotype O1 ATCC 39541 (A0A0H3AMA6 AND A5F1V0 AND A5F5P5), Vibrio cholerae serotype O1 O395 (A0A0H3AMA6 AND A5F1V0 AND A5F5P5)
brenda
Priess, M.; Schaefer, L.V.
Release of entropic spring reveals conformational coupling mechanism in the ABC transporter BtuCD-F
Biophys. J.
110
2407-2418
2016
Escherichia coli (P06609 AND P06611 AND P37028), Escherichia coli
brenda
Santos, J.A.; Rempel, S.; Mous, S.T.; Pereira, C.T.; Ter Beek, J.; de Gier, J.W.; Guskov, A.; Slotboom, D.J.
Functional and structural characterization of an ECF-type ABC transporter for vitamin B12
eLife
7
e35828
2018
Lactobacillus delbrueckii subsp. bulgaricus (Q1GBI8), Lactobacillus delbrueckii subsp. bulgaricus DSM 20081 (Q1GBI8), Lactobacillus delbrueckii subsp. bulgaricus NBRC 13953 (Q1GBI8), Lactobacillus delbrueckii subsp. bulgaricus JCM 1002 (Q1GBI8), Lactobacillus delbrueckii subsp. bulgaricus NCIMB 11778 (Q1GBI8), Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842 (Q1GBI8)
brenda
Schmitt, L.
Vitamin B12 import is all about timing
Nat. Chem. Biol.
14
640-641
2018
Escherichia coli (P06609 AND P06611 AND P37028)
brenda
Goudsmits, J.M.H.; Slotboom, D.J.; van Oijen, A.M.
Single-molecule visualization of conformational changes and substrate transport in the vitamin B12 ABC importer BtuCD-F
Nat. Commun.
8
1652
2017
Escherichia coli (P06609 AND P06611 AND P37028)
brenda
Rempel, S.; Colucci, E.; de Gier, J.W.; Guskov, A.; Slotboom, D.J.
Cysteine-mediated decyanation of vitamin B12 by the predicted membrane transporter BtuM
Nat. Commun.
9
3038
2018
Thiobacillus denitrificans (Q3SFD8)
brenda
Pan, C.; Weng, J.; Wang, W.
ATP hydrolysis induced conformational changes in the vitamin B12 transporter BtuCD revealed by MD simulations
PLoS ONE
11
e0166980
2016
Escherichia coli (P06609 AND P06611 AND P37028)
brenda
Kawaguchi, K.; Okamoto, T.; Morita, M.; Imanaka, T.
Translocation of the ABC transporter ABCD4 from the endoplasmic reticulum to lysosomes requires the escort protein LMBD1
Sci. Rep.
6
30183
2016
Homo sapiens (O14678)
brenda
Mireku, S.A.; Sauer, M.M.; Glockshuber, R.; Locher, K.P.
Structural basis of nanobody-mediated blocking of BtuF, the cognate substrate-binding protein of the Escherichia coli vitamin B12 transporter BtuCD
Sci. Rep.
7
14296
2017
Escherichia coli (P06609 AND P06611 AND P37028), Escherichia coli
brenda
Mireku, S.A.; Ruetz, M.; Zhou, T.; Korkhov, V.M.; Kraeutler, B.; Locher, K.P.
Conformational change of a tryptophan residue in BtuF facilitates binding and transport of cobinamide by the vitamin B12 transporter BtuCD-F
Sci. Rep.
7
41575
2017
Escherichia coli (P06609 AND P06611 AND P37028), Escherichia coli
brenda