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Literature summary extracted from

  • Reyes, A.C.; Amyes, T.L.; Richard, J.P.
    Enzyme architecture: a startling role for Asn270 in glycerol 3-phosphate dehydrogenase-catalyzed hydride transfer (2016), Biochemistry, 55, 1429-1432.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.8 phosphite dianion strongly activates the wild-type enzyme. The N270A mutation results in a change in the effect of phosphite dianion on (kcat/Km)obs for GPDH-catalyzed reduction of glycerone phosphate, from strongly activating to inhibiting Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.8 N270A site-direted mutagenesis, the mutation results in a 7.7 kcal/mol decrease in the intrinsic phosphodianion binding energy, which is larger than the 5.6 kcal/mol effect of the mutation on the stability of the transition state for reduction of DHAP. A 2.2 kcal/mol stabilization of the transition state for unactivated hydride transfer to the truncated substrate glycolaldehyde, and a change in the effect of phosphite dianion on GPDH-catalyzed reduction of glycolaldehyde, from strongly activating to inhibiting. The N270A mutation breaks the network of hydrogen bonding side chains, Asn270, Thr264, Asn205, Lys204, Asp260, and Lys120, which connect the dianion activation and catalytic sites of GPDH. The disruption dramatically alters the performance of GPDH at these sites. The mutant enzyme shows highly decreased activity compared to the wild-type with glycerone phosphate,but 40fold higher activity with glycolaldehyde compared to the wild-type. There is no significant rescue of the activity of the N270A mutant with glycerone phosphate by 60 mM formamide Homo sapiens
1.1.1.8 N270A the mutant shows a strong decrease in catalytic efficiency compared to the wild type enzyme Homo sapiens
1.1.1.8 R269A site-direted mutagenesis, the mutant enzyme shows highly reduced activity compared to the wild-type Homo sapiens
1.1.1.8 R269A the mutant shows a strong decrease in catalytic efficiency compared to the wild type enzyme Homo sapiens
1.1.1.8 R269A/N270A site-direted mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type. The N270A mutation at R269A hlGPDH results in the loss of the intrinsic side chain-dianion interaction Homo sapiens
1.1.1.8 R269A/N270A the mutant shows a strong decrease in catalytic efficiency compared to the wild type enzyme Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.8 phosphite dianion strongly inhibits the mutant N270A enzyme. The N270A mutation results in a change in the effect of phosphite dianion on (kcat/Km)obs for GPDH-catalyzed reduction of glycerone phosphate, from strongly activating to inhibiting Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.8 additional information
-
additional information Michaelis-Menten kinetics Homo sapiens
1.1.1.8 0.052
-
glycerone phosphate wild-type enzyme, pH 7.5, 25°C Homo sapiens
1.1.1.8 0.052
-
dihydroxyacetone phosphate wild type enzyme, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 1.5
-
glycerone phosphate mutant R269A/N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 2.5
-
glycerone phosphate mutant N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 5.7
-
dihydroxyacetone phosphate mutant enzyme R269A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 5.7
-
glycerone phosphate mutant R269A, pH 7.5, 25°C Homo sapiens
1.1.1.8 15
-
dihydroxyacetone phosphate mutant enzyme R269A/N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 25
-
dihydroxyacetone phosphate mutant enzyme N270A, at pH 7.5 and 25°C Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.8 glycerone phosphate + NADH + H+ Homo sapiens
-
sn-glycerol 3-phosphate + NAD+
-
?
1.1.1.8 sn-glycerol 3-phosphate + NAD+ Homo sapiens
-
glycerone phosphate + NADH + H+
-
r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.8 Homo sapiens
-
-
-
1.1.1.8 Homo sapiens P21695
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.8 liver
-
Homo sapiens
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.8 dihydroxyacetone phosphate + NADH + H+
-
Homo sapiens ?
-
?
1.1.1.8 glycerone phosphate + NADH + H+
-
Homo sapiens sn-glycerol 3-phosphate + NAD+
-
?
1.1.1.8 glycerone phosphate + NADH + H+
-
Homo sapiens sn-glycerol 3-phosphate + NAD+
-
r
1.1.1.8 glycolaldehyde + NADH + H+
-
Homo sapiens ?
-
?
1.1.1.8 glycolaldehyde + NADH + H+ truncated substrate Homo sapiens ? + NAD+
-
r
1.1.1.8 sn-glycerol 3-phosphate + NAD+
-
Homo sapiens glycerone phosphate + NADH + H+
-
r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.8 glycerol 3-phosphate dehydrogenase
-
Homo sapiens
1.1.1.8 GPDH
-
Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.8 25
-
assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.8 0.00028
-
dihydroxyacetone phosphate mutant enzyme R269A/N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.00028
-
glycerone phosphate mutant R269A/N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 0.0059
-
dihydroxyacetone phosphate mutant enzyme R269A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.0059
-
glycerone phosphate mutant R269A, pH 7.5, 25°C Homo sapiens
1.1.1.8 9
-
dihydroxyacetone phosphate mutant enzyme N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 9
-
glycerone phosphate mutant N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 240
-
glycerone phosphate wild-type enzyme, pH 7.5, 25°C Homo sapiens
1.1.1.8 240
-
dihydroxyacetone phosphate wild type enzyme, at pH 7.5 and 25°C Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.8 7.5
-
assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.8 NAD+
-
Homo sapiens
1.1.1.8 NADH
-
Homo sapiens

General Information

EC Number General Information Comment Organism
1.1.1.8 malfunction mutation N270A results in a 7.7 kcal/mol decrease in the intrinsic phosphodianion binding energy, which is larger than the 5.6 kcal/mol effect of the mutation on the stability of the transition state for reduction of DHAP. A 2.2 kcal/mol stabilization of the transition state for unactivated hydride transfer to the truncated substrate glycolaldehyde, and a change in the effect of phosphite dianion on GPDH-catalyzed reduction of glycolaldehyde, from strongly activating to inhibiting. The N270A mutation breaks the network of hydrogen bonding side chains, Asn270, Thr264, Asn205, Lys204, Asp260, and Lys120, which connect the dianion activation and catalytic sites of GPDH. The disruption dramatically alters the performance of GPDH at these sites Homo sapiens
1.1.1.8 additional information structure-function analysis of wild-type and mutant enzymes, overview Homo sapiens
1.1.1.8 physiological function role for Asn270 in glycerol 3-phosphate dehydrogenase-catalyzed hydride transfer Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.8 0.000003
-
glycolaldehyde mutant enzyme R269A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.000003
-
glycolaldehyde mutant enzyme R269A/N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.00019
-
glycerone phosphate mutant R269A/N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 0.001
-
dihydroxyacetone phosphate mutant enzyme R269A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.001
-
glycerone phosphate mutant R269A, pH 7.5, 25°C Homo sapiens
1.1.1.8 0.002
-
glycolaldehyde mutant enzyme N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.17
-
dihydroxyacetone phosphate mutant enzyme R269A/N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.36
-
dihydroxyacetone phosphate mutant enzyme N270A, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 0.5
-
glycolaldehyde wild type enzyme, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 3
-
glycolaldehyde mutant R269A, pH 7.5, 25°C Homo sapiens
1.1.1.8 3
-
glycolaldehyde mutant R269A/N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 3.6
-
glycerone phosphate mutant N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 50
-
glycolaldehyde wild-type enzyme, pH 7.5, 25°C Homo sapiens
1.1.1.8 2000
-
glycolaldehyde mutant N270A, pH 7.5, 25°C Homo sapiens
1.1.1.8 4600
-
dihydroxyacetone phosphate wild type enzyme, at pH 7.5 and 25°C Homo sapiens
1.1.1.8 4615
-
glycerone phosphate wild-type enzyme, pH 7.5, 25°C Homo sapiens