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Literature summary for 4.1.2.13 extracted from

  • Pegan, S.D.; Rukseree, K.; Capodagli, G.C.; Baker, E.A.; Krasnykh, O.; Franzblau, S.G.; Mesecar, A.D.
    Active site loop dynamics of a class IIa fructose 1,6-bisphosphate aldolase from Mycobacterium tuberculosis (2013), Biochemistry, 52, 912-925.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with inhibitor phosphoglycolohydroxamate as a mimic of the hydroxyenolate intermediate- and dihydroxyacetone phosphate-bound form of the enzyme, to 1.58 A resolution. Residue E169 facilitates a water-mediated deprotonation-protonation step of the reaction mechanism Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D276A about 1% of wild-type activity Mycobacterium tuberculosis
E168A about 30% of wild-type activity Mycobacterium tuberculosis
E169A almost complete loss of activity Mycobacterium tuberculosis
G167A about 3% of wild-type activity Mycobacterium tuberculosis
G167A/G166A about 2% of wild-type activity Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
dihydroxyacetone phosphate competitive with fructose 1,6-bisphosphate Mycobacterium tuberculosis
Phosphoglycolohydroxamate use as a mimic of the hydroxyenolate intermediate- and dihydroxyacetone phosphate-bound form of the enzyme Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.018
-
D-fructose-1,6-bisphosphate mutant E169A, pH 7.8, 22°C Mycobacterium tuberculosis
0.021
-
D-fructose-1,6-bisphosphate mutant G167A, pH 7.8, 22°C Mycobacterium tuberculosis
0.021
-
D-fructose-1,6-bisphosphate mutant G167A/G166A, pH 7.8, 22°C Mycobacterium tuberculosis
0.03
-
D-fructose-1,6-bisphosphate mutant E168A, pH 7.8, 22°C Mycobacterium tuberculosis
0.04
-
D-fructose-1,6-bisphosphate wild-type, pH 7.8, 22°C Mycobacterium tuberculosis
0.18
-
D-fructose-1,6-bisphosphate mutant D276A, pH 7.8, 22°C Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WQA3
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose-1,6-bisphosphate
-
Mycobacterium tuberculosis dihydroxyacetone phosphate + glyceraldehyde-3-phosphate
-
?
additional information product glycerone phosphate is leaving first followed by dihydroxyacetone phosphate in a uni-bi kinetic scheme Mycobacterium tuberculosis ?
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
D-fructose-1,6-bisphosphate mutant E169A, pH 7.8, 22°C Mycobacterium tuberculosis
0.11
-
D-fructose-1,6-bisphosphate mutant G167A/G166A, pH 7.8, 22°C Mycobacterium tuberculosis
0.17
-
D-fructose-1,6-bisphosphate mutant G167A, pH 7.8, 22°C Mycobacterium tuberculosis
0.45
-
D-fructose-1,6-bisphosphate mutant D276A, pH 7.8, 22°C Mycobacterium tuberculosis
2.67
-
D-fructose-1,6-bisphosphate mutant E168A, pH 7.8, 22°C Mycobacterium tuberculosis
12
-
D-fructose-1,6-bisphosphate wild-type, pH 7.8, 22°C Mycobacterium tuberculosis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.238
-
dihydroxyacetone phosphate wild-type, pH 7.8, 22°C Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.28
-
D-fructose-1,6-bisphosphate mutant E169A, pH 7.8, 22°C Mycobacterium tuberculosis
2.5
-
D-fructose-1,6-bisphosphate mutant D276A, pH 7.8, 22°C Mycobacterium tuberculosis
5.33
-
D-fructose-1,6-bisphosphate mutant G167A/G166A, pH 7.8, 22°C Mycobacterium tuberculosis
8
-
D-fructose-1,6-bisphosphate mutant G167A, pH 7.8, 22°C Mycobacterium tuberculosis
88.33
-
D-fructose-1,6-bisphosphate mutant E168A, pH 7.8, 22°C Mycobacterium tuberculosis
300
-
D-fructose-1,6-bisphosphate wild-type, pH 7.8, 22°C Mycobacterium tuberculosis