Cloned (Comment) | Organism |
---|---|
recombinant expression of N-terminal His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS Rosetta | Streptococcus agalactiae |
Crystallization (Comment) | Organism |
---|---|
purified His-tagged enzyme in complex with NAD+, microseeding method, mixing of 8.9 mg/ml protein in 25 mM HEPES, pH 7.35, 0.1 M NaCl, and 5 mM 2-mercaptoethanol, with crystallization solution containing 26-36% PEG 4000, 0.1 M MES, pH 6.5, the needle-like crystals are used for microseeding by mixing 0.002 ml of protein plus cofactor mixture, 500 nl water and 500 nl reservoir solution containing seed suspension in 28% PEG 4000, 0.1 M MES, pH 6.5, X-ray diffraction structure determination and analysis at 2.46 A resolution, molecular replacement and modelling | Streptococcus agalactiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Streptococcus agalactiae | 5829 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | Streptococcus agalactiae | - |
3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | Streptococcus agalactiae NEM316 | - |
3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NADP+ | Streptococcus agalactiae | - |
3-phospho-D-glyceroyl phosphate + NADPH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NADP+ | Streptococcus agalactiae NEM316 | - |
3-phospho-D-glyceroyl phosphate + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus agalactiae | Q8E3E8 | - |
- |
Streptococcus agalactiae NEM316 | Q8E3E8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminal His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS Rosetta by nickel affinity chromatography, ultrafiltration, and gel filtration | Streptococcus agalactiae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD(P)+ = 3-phospho-D-glyceroyl phosphate + NAD(P)H + H+ | GAPDH catalyzes the oxidative phosphorylation of D-glyceraldehyde 3-phosphate (D-G3H) to give 1,3-diphosphoglyceric acid and uses NAD+ or NADP+ as a cofactor. In the first step of this two-step reaction the active-site cysteine residue of GAPDH attaches covalently to D-G3H and forms a thiohemiacetal intermediate, which transfers a hydride ion to NAD+, resulting in the formation of the thioacyl enzyme. In the second step the resulting thioester is phosphorylated through the nucleophilic attack of an inorganic phosphate ion on the carbonyl C atom of the thioacyl group, which leads to the formation of 1,3-diphosphoglycerate | Streptococcus agalactiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Streptococcus agalactiae | 3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Streptococcus agalactiae NEM316 | 3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NADP+ | - |
Streptococcus agalactiae | 3-phospho-D-glyceroyl phosphate + NADPH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NADP+ | - |
Streptococcus agalactiae NEM316 | 3-phospho-D-glyceroyl phosphate + NADPH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | crystal structure analysis | Streptococcus agalactiae |
Synonyms | Comment | Organism |
---|---|---|
GAPDH | - |
Streptococcus agalactiae |
GBS GAPDH | - |
Streptococcus agalactiae |
glyceraldehyde-3-phosphate dehydrogenase | - |
Streptococcus agalactiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | GAPDH catalyzes the oxidative phosphorylation of D-glyceraldehyde 3-phosphate using NAD+ or NADP+ as a cofactor | Streptococcus agalactiae | |
NAD+ | enzyme binding structure analysis | Streptococcus agalactiae | |
NADP+ | - |
Streptococcus agalactiae |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme plays a key role in glycolysis. GAPDH localized on the surface of some bacteria is thought to be involved in macromolecular interactions and bacterial pathogenesis. GAPDH on the surface of group B streptococcus (GBS) enhances bacterial virulence and is a potential vaccine candidate | Streptococcus agalactiae |