Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain HB101 | Homo sapiens |
recombinant expression of wild-type enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain HB101 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D311N | site-directed mutagenesis, the mutation breaks the salt bridge between the catalytic and NAD+-binding domains, mutant dN-GAPDS D311N binds NAD+ noncooperatively | Homo sapiens |
E244Q | site-directed mutagenesis, mutation at the interdomain salt bridge, the E244Q substitution does not alter the NAD+-binding significantly. The mutant protein exhibits a well-pronounced positive cooperativity in coenzyme binding | Homo sapiens |
E96Q | site-directed mutagenesis, mutation at the interdomain salt bridge, the E96Q substitution does not alter the NAD+-binding significantly. The mutant protein exhibits a well-pronounced positive cooperativity in coenzyme binding | Homo sapiens |
additional information | construction of a plasmid encoding truncated GAPDS lacking 68 N-terminal amino acids (dN-GAPDS). The recombinant GAPDS without the N-terminal sequence (dN-GAPDS) is soluble in contrast to the wild-type | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics, overview | Homo sapiens | |
additional information | - |
additional information | the sperm-specific isoenzyme of glyceraldehyde-3-phosphate dehydrogenase exhibits strong positive cooperativity in coenzyme binding, kinetics, overview | Homo sapiens | |
0.097 | - |
D-glyceraldehyde 3-phosphate | recombinant dN-GAPDS enzyme, pH 8.9, 20°C | Homo sapiens | |
0.24 | - |
D-glyceraldehyde 3-phosphate | recombinant dN-GAPDS enzyme mutant D311N, pH 8.9, 20°C | Homo sapiens | |
0.27 | - |
D-glyceraldehyde 3-phosphate | recombinant wild-type enzyme, pH 8.9, 20°C | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
flagellum | GAPDS is an insoluble protein that is attached to the cytoskeleton of the sperm flagellum via the N-terminus | Homo sapiens | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
150000 | - |
native PAGE | Homo sapiens |
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+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | O14556 | - |
- |
Homo sapiens | P04406 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and truncated mutant dN-GAPDS by cation exchange chromatography from Escherichia coli strain BL21(DE3), mutant dNGAPDS D311N by ammonium sulfate fractionation, and dialysis, followed by anion exchange chromatography, the flow-through is further purified by hydrophobic interaction chromatography | Homo sapiens |
recombinant wild-type enzyme from Escherichia coli strain BL21(DE3) | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
skeletal muscle | - |
Homo sapiens | - |
sperm cell | - |
Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
45 | - |
purified recombinant truncated enzyme mutant dN-GAPDS, pH 8.9, 20°C, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
60 | - |
purified recombinant truncated enzyme mutant dN-GAPDS E244Q, pH 8.9, 20°C, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
64 | - |
purified recombinant truncated enzyme mutant dN-GAPDS E96Q, pH 8.9, 20°C, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
92 | - |
purified recombinant truncated enzyme mutant dN-GAPDS D311N, pH 8.9, 20°C, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
95 | - |
purified recombinant wild-type enzyme, pH 8.9, 20°C, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Homo sapiens | 3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 37000, about, SDS-PAGE | Homo sapiens |
More | three GAPDS-specific salt bridges, E96-H394 and D311-H124 connect the NAD+-binding and the catalytic domains within a single subunit, while E244-R320 is formed between two different subunits | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
GAPDS | - |
Homo sapiens |
glyceraldehyde-3-phosphate dehydrogenase | - |
Homo sapiens |
somatic GAPD | - |
Homo sapiens |
sperm-specific GAPDS | - |
Homo sapiens |
sperm-specific glyceraldehyde-3-phosphate dehydrogenase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | - |
assay at, oxidation | Homo sapiens |
20 | - |
assay at, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.9 | - |
assay at, oxidation | Homo sapiens |
8.9 | - |
assay at, oxidation of D-glyceraldehyde 3-phosphate | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | monomer interactions with all three neighbor subunits are essential for maintaining the cooperativity of NAD+ binding exhibited by GAPD, NAD+ binding analysis, kinetics, overview | Homo sapiens | |
NAD+ | NAD+ binding analysis with truncated and mutant enzymes, kinetics, overview. The enzyme exhibits positive cooperativity with the saturation being reached at the NAD+/protein concentration ratio of about 20. The E96Q and E244Q substitutions do not alter the NAD+-binding behavior of dN-GAPDS significantly. The mutant proteins exhibit a well-pronounced positive cooperativity in coenzyme binding, while mutant dN-GAPDS D311N binds NAD+ noncooperatively | Homo sapiens | |
NADH | - |
Homo sapiens |