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 (Comment) | Organism |
---|---|
gene gapA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Escherichia coli |
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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9B2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltratioon | Escherichia coli |
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 | 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 | Comment | Organism |
---|---|---|
EcGAPDH1 | - |
Escherichia coli |
GapA | - |
Escherichia coli |
GAPDH type 1 | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Escherichia coli |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 60 | over 50% of maximal activity within this range, profile overview | Escherichia coli |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10 | - |
- |
Escherichia coli |
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 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 | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Escherichia coli |
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 |