Application | Comment | Organism |
---|---|---|
pharmacology | the inhibition by lithium suggests the FIG superfamily of enzymes is the target of lithium therapy in manic-depressive illness | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
gene cysQ, sequence comparisons, overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant Mycobacterium tuberculosis enzyme CysQ in a ligand-free structure, a lithium-inhibited state with substrate PAP bound, and a product-bound complex with AMP, phosphate, and three Mg2+ ions bound, sitting drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris, pH 8.0, 100 mM NaCl, 1 mM DTT, 5% glycerol, and 1 mM AMP, with 0.001 ml of reservoir solution containing 24% PEG 1500 and 20% glycerol for the ligand-free enzyme, the calcium- and phosphate-bound structure is grown in 0.05 M CaCl2, 0.1 M Bis-Tris-HCl, pH 6.5, and 30% PEG MME 550, the magnesium- and phosphate-bound structure is grown in 0.1 M Mg acetate, 0.1 M Bis-Tris-HCl, pH 6.5, and 35% PEG MME 550, and for both the Ca2+ and Mg2+ structures, phosphate is not included in the crystallization conditions, the AMP-, Pi-, and Mg2+-bound crystals are grown in 0.1 M Mg acetate, 0.1 M Bis-Tris-HCl, pH 6.5, and 35% PEG MME 550 and soaked for 20 min in mother liquor supplemented with 3 mM AMP, 21°C, 1-5 days, X-ray diffraction structure determination and analysis at 1.45-2.05 A resolution. SIRAS (single isomorphous replacement with anomalous scattering) phasing information is collected from a ligand-free crystal grown in 0.2 M calcium acetate, 0.1 M MES-NaOH, pH 6.0, and 20% w/v PEG 8000 that is soaked overnight in mother liquor supplemented with a heavy atom cocktail: 1 mM uranyl acetate, 1 mM p-chloromercury benzoic acid sulfonate (PCMBS), and 1 mM KAu(CN)2. Before being flash-cooled, the crystal is transferred to a cryosolution containing 0.2 M Ca-acetate, 0.1 M MES-NaOH, pH 6.0, 20% w/v PEG 8000, and 20% glycerol. Molecular replacement and structure modelling | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Li+ | the Li+ ion displaces Mg2+ ion, specifically the Mg2+ ion at metal site 2, and is thought to stabilize the phosphate-bound product. Enzyme binding structure, overview | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates, enzyme binding structure, overview | Mycobacterium tuberculosis | |
Mg2+ | activates, three Mg2+ ions binding in the active site are required to form a productive enzyme, enzyme binding structure, overview | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
adenosine 3',5'-bisphosphate + H2O | Mycobacterium tuberculosis | - |
AMP + phosphate | - |
? | |
adenosine 3',5'-bisphosphate + H2O | Mycobacterium tuberculosis H37Rv | - |
AMP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WKJ1 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WKJ1 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and ultrafiltration | Mycobacterium tuberculosis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
adenosine 3',5'-bisphosphate + H2O = AMP + phosphate | proposed catalytic mechanism for CysQ, Asp50 deprotonates Thr96 that deprotonates the water (ligating a Mg2+), which attacks the phosphate, the pentacovalent 3'-phosphate transition state is stabilized by the three Mg2+ ions, overview | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
adenosine 3',5'-bisphosphate + H2O | - |
Mycobacterium tuberculosis | AMP + phosphate | - |
? | |
adenosine 3',5'-bisphosphate + H2O | i.e. PAP | Mycobacterium tuberculosis | AMP + phosphate | - |
? | |
adenosine 3',5'-bisphosphate + H2O | - |
Mycobacterium tuberculosis H37Rv | AMP + phosphate | - |
? | |
adenosine 3',5'-bisphosphate + H2O | i.e. PAP | Mycobacterium tuberculosis H37Rv | AMP + phosphate | - |
? | |
additional information | Mtb CysQ can dephosphorylate myo-inositol 1-phosphate (IMP), fructose 1,6-bisphosphate (FBP), and 3'-phosphoadenosine 5'-monophosphate (PAP), but the catalytic efficiency is 1700 and 15000fold greater for PAP over FBP and IMP, respectively. AMP and phosphate enzyme binding structures, overview. The specificity of a 5'-phosphate or bis-phosphate sugars for CysQ can likely come from a conserved positively charged residue, Lys190 in CysQ, that ion pairs with the nonhydrolyzed phosphate | Mycobacterium tuberculosis | ? | - |
? | |
additional information | Mtb CysQ can dephosphorylate myo-inositol 1-phosphate (IMP), fructose 1,6-bisphosphate (FBP), and 3'-phosphoadenosine 5'-monophosphate (PAP), but the catalytic efficiency is 1700 and 15000fold greater for PAP over FBP and IMP, respectively. AMP and phosphate enzyme binding structures, overview. The specificity of a 5'-phosphate or bis-phosphate sugars for CysQ can likely come from a conserved positively charged residue, Lys190 in CysQ, that ion pairs with the nonhydrolyzed phosphate | Mycobacterium tuberculosis H37Rv | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3',5'-bisphosphate nucleotidase | - |
Mycobacterium tuberculosis |
CysQ | - |
Mycobacterium tuberculosis |
MTCY270.37 | - |
Mycobacterium tuberculosis |
PAP phosphatase | - |
Mycobacterium tuberculosis |
Rv2131c | - |
Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
evolution | CysQ is part of the larger FIG superfamily of phosphatases that dephosphorylates a monosaccharide-containing substrate. The FIG superfamily is comprised of family members: fructose-1,6-bisphosphatase (FBPase), inositol-monophosphatases (IMPase)/polyphosphatases (IPPase), and the glpX-encoded variant of FBPase (Class II). Many members of this superfamily display promiscuous phosphatase activity toward various monosaccharide-containing substrates but are usually more efficient in one type, which identifies their subfamily class. Mtb CysQ can dephosphorylate myo-inositol 1-phosphate (IMP), fructose 1,6-bisphosphate (FBP), and 3'-phosphoadenosine 5'-monophosphate (PAP), but the catalytic efficiency is 1700 and 15000fold greater for PAP over FBP and IMP, respectively. The FIG superfamily enzymes have demonstrated a dependence on divalent metal ions and are most active with magnesium. The superfamily also displays sensitivity to monovalent metals, and members are most strongly inhibited by lithium (Li+) | Mycobacterium tuberculosis |
malfunction | cysQ knockout results in reduced levels of the sulfated glycolipid sulfolipid-1 and attenuation of cell growth | Mycobacterium tuberculosis |
physiological function | because excess 3'-phosphoadenosine 5'-phosphate (PAP) alters the equilibrium of the sulfur pathway and inhibits sulfotransferases, PAP concentrations can affect the levels of sulfur-containing metabolites. PAP is removed by the phosphatase activity of CysQ, a 3',5'-bisphosphate nucleotidase, yielding AMP and phosphate, CysQ, a divalent cation metal-dependent phosphatase, is a major regulator of the sulfur activation pathway | Mycobacterium tuberculosis |