Literature summary for 1.2.1.59 extracted from

Ayres, C.A.; Schormann, N.; Senkovich, O.; Fry, A.; Banerjee, S.; Ulett, G.C.; Chattopadhyay, D.
Structure of Streptococcus agalactiae glyceraldehyde-3-phosphate dehydrogenase holoenzyme reveals a novel surface (2014), Acta Crystallogr. Sect. F, 70, 1333-1339 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminal His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS Rosetta	Streptococcus agalactiae

Crystallization (Commentary)

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

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

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Streptococcus agalactiae	5829	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
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

Organism	UniProt	Comment	Textmining
Streptococcus agalactiae	Q8E3E8	-	-
Streptococcus agalactiae NEM316	Q8E3E8	-	-

Purification (Commentary)

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

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	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Subunits	Comment	Organism
tetramer	crystal structure analysis	Streptococcus agalactiae

Synonyms

Synonyms	Comment	Organism
GAPDH	-	Streptococcus agalactiae
GBS GAPDH	-	Streptococcus agalactiae
glyceraldehyde-3-phosphate dehydrogenase	-	Streptococcus agalactiae

Cofactor

Cofactor	Comment	Organism
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

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