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Literature summary for 1.2.1.12 extracted from

  • Zhang, L.; Liu, M.; Yao, Y.; Bostrom, I.; Wang, Y.; Chen, A.; Li, J.; Gu, S.; Ji, C.
    Characterization and structure of glyceraldehyde-3-phosphate dehydrogenase type 1 from Escherichia coli (2020), Acta Crystallogr. Sect. F, 76, 406-413 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information no effect on enzyme activity by SDS at 1% Escherichia coli
Triton X-100 activates 30% at 1% Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
gene gapA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant isozyme EcGAPDH1, sitting drop vapour diffusion method, mixing of 240 nl of 30 mg/ml protein in 4 mM NaCl, and 5 mM Tris-HCl pH 8.0, with 240 nl of reservoir solution containing reservoir solution consisting of 100 mM sodium acetate, pH 4.6, 30% w/v PEG 400,and 200 mM calcium acetate, and equilibration against 0.1 ml of reservoir solution, 1 week, method optimization, X-ray diffraction structure determination and analysis at 1.88 A resolution, molecular replacement using the structure of GAPDH from methicillin-resistant Staphylococcus aureus MRSA252 (PDB ID 3lvf) as search model, model building Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
acetone inhibits about 60% at 15% Escherichia coli
acetonitril inhibits about 90% at 15% Escherichia coli
ethanol inhibits about 55% at 15% Escherichia coli
Isopropanol inhibits about 60% at 15% Escherichia coli
additional information no effect on enzyme activity by SDS at 1% Escherichia coli
Tween 20 inhibits about 60% at 1% Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+ Escherichia coli
-
3-phospho-D-glyceroyl phosphate + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A9B2
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltratioon Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
Escherichia coli 3-phospho-D-glyceroyl phosphate + NADH + H+
-
?

Subunits

Subunits Comment Organism
tetramer each of the subunits can be divided into two domains: the N-terminal NAD+-binding domain and the C-terminal catalytic domain. The NAD+-binding domain is typically a Rossman fold containing eight beta-strands, namely beta1 (Lys3-Asn7), beta2 (Asp28-Asn33), beta3 (Val58-Phe60), beta4 (Ser64-Val67), beta5 (Lys70-Tyr75), beta6 (Ile92-Glu95), beta7 (Lys116-Ile119) and beta8 (Ile144-Ser146). The strands are connected by either helices or short loops. beta3 and beta5 are antiparallel to the other six parallel beta-strands. There are four alpha-helices in this domain: alpha1 (Gly10-Val23), alpha2 (Ser37-His47), alpha3 (Ser102-Ser106) and alpha4 (Gln107-Ala112). The catalytic domain contains eight mixed beta-sheets, beta9 (Ile168-Ala178), beta10 (Ile205-His207), beta11 (Leu226-Val231), beta12 (Ser239-Leu247), beta13 (Phe270-Thr273), beta14 (Ser289-Asp292), beta15 (Glu297-Val301) and beta16 (Leu304-Tyr313), and three long alpha-helices, alpha5 (Ser149-Gly167), alpha6 (Thr252-Thr264) and alpha7 (Gln317-Lys332). The catalytically active residues Cys150 and His177 are situated in alpha5 and beta9, respectively Escherichia coli

Synonyms

Synonyms Comment Organism
EcGAPDH1
-
Escherichia coli
GapA
-
Escherichia coli
GAPDH type 1
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Escherichia coli

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
45 60 over 50% of maximal activity within this range, profile overview Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
purified recombinant His-tagged enzyme, completely stable up to Escherichia coli
55
-
purified recombinant His-tagged enzyme, loss of 70-80% activity Escherichia coli
65
-
purified recombinant His-tagged enzyme, inactivation Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
-
Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
8 10.5 over 40% of maximal activity within this range, profile overview Escherichia coli

pH Stability

pH Stability pH Stability Maximum Comment Organism
1 2 purified recombinant His-tagged enzyme, loss of 80% activity Escherichia coli
7 10 purified recombinant His-tagged enzyme, completely stable Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Escherichia coli

General Information

General Information Comment Organism
metabolism glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic pathway that catalyzes the conversion of D-glyceraldehyde 3-phosphate to 1,3-diphosphoglycerate Escherichia coli
additional information three-dimensional structure analysis of EcGAPDH1 compared with the structures of HuGAPDH and MrsaGAPDH shows that the main difference is the loop conformation, especially the S-loop Escherichia coli
physiological function glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic pathway that catalyzes the conversion of D-glyceraldehyde 3-phosphate to 1,3-diphosphoglycerate Escherichia coli