Literature summary for 3.1.3.11 extracted from

Wolf, N.; Gutka, H.; Movahedzadeh, F.; Abad-Zapatero, C.
Structures of the mycobacterium tuberculosis glpx protein (Class II fructose-1,6-bisphosphatase) Implications for the active oligomeric state, catalytic mechanism and citrate inhibition (2018), Acta Crystallogr. Sect. D, 74, 321-331 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His-tagged wild-type and mutant enzymes	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging-drop vapor-diffusion method at 25°C, crystal structures of native enzyme (FBPaseII) at 2.6 A resolution and two active-site protein variants. The variants are complexed with the reaction product fructose 6-phosphate. Presence of a 222 tetramer	Mycobacterium tuberculosis
purified recombinant wild-type apoform and two active site mutants T84A and T84S in complex with D-fructose 1,6-bisphosphate or D-fructose 6-phosphate (F6P), hanging drop vapor diffusion method, mixing of protein and precipitant solution in a 1:1 ratio, the reservoir solution contains 1.0 M ammonium citrate tribasic, pH 7.0, and 1% PEG 3350, for the apoenzyme, and 2.4 M sodium malonate, pH 6.0, with 1 mM ligand, for the complexed mutant enzyme crystals, X-ray diffraction structure determination and analysis at 2.6 A, 2.3 A, and 2.2 A resolution, respectively, crystal structures of apo MtFBPaseII, the T84A MtFBPase-F6P complex and the T84S MtFBPase-F6P complex are solved by molecular replacement	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

hanging-drop vapor-diffusion method at 25°C, crystal structures of native enzyme (FBPaseII) at 2.6 A resolution and two active-site protein variants. The variants are complexed with the reaction product fructose 6-phosphate. Presence of a 222 tetramer

Mycobacterium tuberculosis

purified recombinant wild-type apoform and two active site mutants T84A and T84S in complex with D-fructose 1,6-bisphosphate or D-fructose 6-phosphate (F6P), hanging drop vapor diffusion method, mixing of protein and precipitant solution in a 1:1 ratio, the reservoir solution contains 1.0 M ammonium citrate tribasic, pH 7.0, and 1% PEG 3350, for the apoenzyme, and 2.4 M sodium malonate, pH 6.0, with 1 mM ligand, for the complexed mutant enzyme crystals, X-ray diffraction structure determination and analysis at 2.6 A, 2.3 A, and 2.2 A resolution, respectively, crystal structures of apo MtFBPaseII, the T84A MtFBPase-F6P complex and the T84S MtFBPase-F6P complex are solved by molecular replacement

Mycobacterium tuberculosis

Protein Variants

Protein Variants	Comment	Organism
T84A	inactive mutant enzyme	Mycobacterium tuberculosis
T84A	site-directed mutagenesis, catalytically inactive active-site mutant	Mycobacterium tuberculosis
T84S	site-directed mutagenesis, active-site mutant, the mutant shows reduced activity compared to wild-type. One Mg2+ ion is found in T84S MtFBPaseII, coordinated to a glycerol molecule and to Asp79, Asp82, and Glu208 near the cleaved 1-phosphate group of the substrate	Mycobacterium tuberculosis
T84S	lower catalytic activity than wild-type enzyme	Mycobacterium tuberculosis

Inhibitors

Inhibitors	Comment	Organism
2-oxoglutarate	-	Mycobacterium tuberculosis
alpha-ketoglutarate	-	Mycobacterium tuberculosis
citrate	-	Mycobacterium tuberculosis
isocitrate	-	Mycobacterium tuberculosis
malate	-	Mycobacterium tuberculosis
malonate	-	Mycobacterium tuberculosis
additional information	no inhibition by isocitrate	Mycobacterium tuberculosis
oxaloacetate	-	Mycobacterium tuberculosis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required, enzyme-bound	Mycobacterium tuberculosis
additional information	the apoenzyme does not contain Mg2+ in the active site, instead Mg2+ is found in helices H8 and H9 in one chain, coordinated to Leu165 N, Gln164 N and Arg161 O and possibly stabilizing this loop. The fructose 6-phosphate-bound structures contain one magnesium ion in a position that is proposed to bind fructose 1,6-bisphosphate, coordinate the 1-phosphate and stabilize the transition state	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-fructose 1,6-bisphosphate + H2O	Mycobacterium tuberculosis	-	D-fructose 6-phosphate + phosphate	-	?
D-fructose 1,6-bisphosphate + H2O	Mycobacterium tuberculosis ATCC 25618	-	D-fructose 6-phosphate + phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WN20	-	-
Mycobacterium tuberculosis	P9WN21	-	-
Mycobacterium tuberculosis ATCC 25618	P9WN21	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Mycobacterium tuberculosis
recombinant N-terminally His-tagged wild-type and mutant enzymes by nickel affinity chromatography and gel filtration	Mycobacterium tuberculosis

Reaction

Reaction	Comment	Organism	Reaction ID
D-fructose 1,6-bisphosphate + H2O = D-fructose 6-phosphate + phosphate	the proposed mechanism involves two metal ions near the cleavable phosphate: one to stabilize the negative charge on the leaving group and one to coordinate the nucleophilic water	Mycobacterium tuberculosis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-fructose 1,6-bisphosphate + H2O	-	Mycobacterium tuberculosis	D-fructose 6-phosphate + phosphate	-	?
D-fructose 1,6-bisphosphate + H2O	-	Mycobacterium tuberculosis ATCC 25618	D-fructose 6-phosphate + phosphate	-	?

Subunits

Subunits	Comment	Organism
homotetramer	-	Mycobacterium tuberculosis
homotetramer	a 222 tetramer	Mycobacterium tuberculosis
More	the constellation of amino-acid residues in the active site of FBPaseII from Mycobacterium tuberculosis (MtFBPaseII) is conserved and is analogous to that described previously for the Escherichia coli enzyme, structure analysis and comparisons, overview	Mycobacterium tuberculosis

Synonyms

Synonyms	Comment	Organism
class II fructose-1,6-bisphosphatase	-	Mycobacterium tuberculosis
FBPaseII	-	Mycobacterium tuberculosis
GlpX protein	-	Mycobacterium tuberculosis
MtFBPaseII	-	Mycobacterium tuberculosis

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
2	-	pH and temperature not specified in the publication	Mycobacterium tuberculosis	citrate
2	-	versus MtFBPaseII, pH and temperature not specified in the publication	Mycobacterium tuberculosis	citrate
3.1	-	pH and temperature not specified in the publication	Mycobacterium tuberculosis	alpha-ketoglutarate
3.1	-	versus MtFBPaseII, pH and temperature not specified in the publication	Mycobacterium tuberculosis	2-oxoglutarate
3.2	-	pH and temperature not specified in the publication	Mycobacterium tuberculosis	malonate
3.2	-	versus MtFBPaseII, pH and temperature not specified in the publication	Mycobacterium tuberculosis	malonate
3.3	-	pH and temperature not specified in the publication	Mycobacterium tuberculosis	malate
3.3	-	versus MtFBPaseII, pH and temperature not specified in the publication	Mycobacterium tuberculosis	malate
6.4	-	pH and temperature not specified in the publication	Mycobacterium tuberculosis	oxaloacetate
6.4	-	versus MtFBPaseII, pH and temperature not specified in the publication	Mycobacterium tuberculosis	oxaloacetate

General Information

General Information	Comment	Organism
evolution	structural comparison of class I versus class II FBPases	Mycobacterium tuberculosis