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Literature summary extracted from

  • Kim, Y.J.
    A cryoprotectant induces conformational change in glyceraldehyde-3-phosphate dehydrogenase (2018), Acta Crystallogr. Sect. F, 74, 277-282 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.12 gene gapA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.2.1.12 purified recombinant enzyme GAPDH complexed with trehalose, hanging drop vapour diffusion method, mixing of 20 mg/ml protein in 20 mM Tris, 130 mM NaCl pH 7.5, with reservoir solution containing 2.8 M ammonium sulfate, 0.1 M MES, pH 5.5-6.5, 4°C, two to three weeks, GAPDH crystals are treated with a cryoprotectant consisting of 15% v/v trehalose at -173°C, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement using the Escherichia coli GAPDH structure (PDB ID 1s7c) as a starting model Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.2.1.12 trehalose might be an inhibitor, trehalose induces a conformational change in ecGAPDH in the current structure. The rotation of GAPDH also induced a conformational change in its active site. This suggests that the binding of trehalose to GAPDH induced a conformational change in its active site to prevent the binding of NAD+, although the NAD+- and trehalose-binding sites differ from one another Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.12 D-glyceraldehyde 3-phosphate + phosphate + NAD+ Escherichia coli
-
 3-phospho-D-glyceroyl phosphate + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.12 Escherichia coli P0A9B2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.12 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and ultrafiltration Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.12 D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
 Escherichia coli 3-phospho-D-glyceroyl phosphate + NADH + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.2.1.12 EcGAPDH
-
 Escherichia coli
1.2.1.12 GAPDH
-
 Escherichia coli
1.2.1.12 glyceraldehyde-3-phosphate dehydrogenase
-
 Escherichia coli

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.2.1.12 22
-
 purified recombinant ecGAPDH is stable at room temperature Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.12 NAD+
-
 Escherichia coli

General Information

EC Number General Information Comment Organism
1.2.1.12 additional information the S-loop of GAPDH is required for interaction of the enzyme with its cofactor and with other proteins. NAD+-bound GAPDH S-loop fixation occurs by the formation of a complex with the coenzyme NAD+. The structure of trehalose-bound ecGAPDH is compared with the structures of both NAD+-free and NAD+-bound ecGAPDH. At the S-loop, the bound trehalose in the GAPDH structure induces a 2.4° rotation compared with the NAD+-free ecGAPDH structure and a 3.1° rotation compared with the NAD+-bound ecGAPDH structure Escherichia coli
1.2.1.12 physiological function glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyses the conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate using NAD+ as a cofactor. It is a moonlighting enzyme playing multiple roles in the regulation of mRNA stability, intracellular membrane trafficking, iron uptake and transport, DNA replication and repair, and nuclear RNA transport Escherichia coli