Literature summary for 1.2.1.12 extracted from

Chaikuad, A.; Shafqat, N.; Al-Mokhtar, R.; Cameron, G.; Clarke, A.R.; Brady, R.L.; Oppermann, U.; Frayne, J.; Yue, W.W.
Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, GAPDS (2011), Biochem. J., 435, 401-409.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3)-R3 cells	Homo sapiens
expression in Escherichia coli	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystallization of a a highly soluble form of GAPDS truncated at the N-terminus, amino acids 69398. The structure in complex with NAD+ and phosphate maps the two anion-recognition sites within the catalytic pocket that correspond to the conserved Ps site and Pi site identified in other organisms. The structure in complex with NAD+ and glycerol shows serendipitous binding of glycerol at the Ps and Pi sites	Homo sapiens
in complex with NAD+ and phosphate, hanging drop vapor diffusion method, using 20% (w/v) poly(ethylene glycol) 3350, 0.2 M sodium/potassium phosphate and 10% (v/v) ethylene glycol or in complex with NAD+ and glycerol, hanging drop vapor diffusion method, 20% PEG 3350, 0.2 M Na2SO4, 10% (v/v) ethylene glycol and 0.1 M Bis-Tris propane (pH 6.5)	Homo sapiens

Crystallization (Comment)

Organism

crystallization of a a highly soluble form of GAPDS truncated at the N-terminus, amino acids 69398. The structure in complex with NAD+ and phosphate maps the two anion-recognition sites within the catalytic pocket that correspond to the conserved Ps site and Pi site identified in other organisms. The structure in complex with NAD+ and glycerol shows serendipitous binding of glycerol at the Ps and Pi sites

Homo sapiens

in complex with NAD+ and phosphate, hanging drop vapor diffusion method, using 20% (w/v) poly(ethylene glycol) 3350, 0.2 M sodium/potassium phosphate and 10% (v/v) ethylene glycol or in complex with NAD+ and glycerol, hanging drop vapor diffusion method, 20% PEG 3350, 0.2 M Na2SO4, 10% (v/v) ethylene glycol and 0.1 M Bis-Tris propane (pH 6.5)

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
additional information	expression of a highly soluble form of GAPDS truncated at the N-terminus, amino acids 69398. Mutant displays a 3fold increase in catalytic efficiency and shows homotetrameric structure	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00025	-	D-glyceraldehyde 3-phosphate	wild-type, pH 8.5, 25°C	Homo sapiens
0.00046	-	D-glyceraldehyde 3-phosphate	N-terminally truncated mutant, pH 8.5, 25°C	Homo sapiens
0.035	-	NAD+	N-terminally truncated mutant, pH 8.5, 25°C	Homo sapiens
0.1	-	NAD+	wild-type, pH 8.5, 25°C	Homo sapiens
0.25	-	D-glyceraldehyde 3-phosphate	glyceraldehyde-3-phosphate dehydrogenase, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens
0.46	-	D-glyceraldehyde 3-phosphate	recombinant, highly soluble form of sperm-specific glyceraldehyde-3-phosphate dehydrogenase truncated at the N-terminus, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens
35	-	NAD+	recombinant, highly soluble form of sperm-specific glyceraldehyde-3-phosphate dehydrogenase truncated at the N-terminus, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens
100	-	NAD+	glyceraldehyde-3-phosphate dehydrogenase, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
37672	-	4 * 37672, MALDI-TOF mass spectrometry	Homo sapiens
38000	-	4 * 38000, SDS-PAGE	Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O14556	-	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-Sepharose column chromatography and Superdex S75 gel filtration	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
sperm	-	Homo sapiens	-

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
homotetramer	4 * 38000, SDS-PAGE	Homo sapiens
homotetramer	4 * 37672, MALDI-TOF mass spectrometry	Homo sapiens
tetramer	N-terminally truncated mutant, crystallization data	Homo sapiens

Synonyms

Synonyms	Comment	Organism
GAPDS	-	Homo sapiens
sperm-specific glyceraldehyde-3-phosphate dehydrogenase	-	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
199	-	NAD+	wild-type, pH 8.5, 25°C	Homo sapiens
199	-	D-glyceraldehyde 3-phosphate	glyceraldehyde-3-phosphate dehydrogenase, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens
234	-	D-glyceraldehyde 3-phosphate	recombinant, highly soluble form of sperm-specific glyceraldehyde-3-phosphate dehydrogenase truncated at the N-terminus, in 10 mM sodium diphosphate, 20 mM sodium phosphate (pH 8.5), 0.003 mM dithiothreitol and 10 mM sodium arsenate, at 25°C	Homo sapiens
234	-	NAD+	N-terminally truncated mutant, pH 8.5, 25°C	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Homo sapiens