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Crystal Structure of Two Ternary Complexes of Phosphorylating Glyceraldehyde-3-phosphate Dehydrogenase from Bacillus stearothermophilus with NAD and d-Glyceraldehyde 3-Phosphate

Abstract : The crystal structure of the phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus was solved in complex with its cofactor, NAD, and its physiological substrate,D-glyceraldehyde 3-phosphate (D-G3P). To isolate a stable ternary complex, the nucleophilic residue of the active site, Cys149, was substituted with alanine or serine. The C149A and C149S GAPDH ternary complexes were obtained by soaking the crystals of the corresponding binary complexes (enzyme·NAD) in a solution containing G3P. The structures of the two binary and the two ternary complexes are presented. The D-G3P adopts the same conformation in the two ternary complexes. It is bound in a non-covalent way, in the free aldehyde form, its C-3 phosphate group being positioned in the Ps site and not in the Pi site. Its C-1 carbonyl oxygen points toward the essential His176, which supports the role proposed for this residue along the two steps of the catalytic pathway. Arguments are provided that the structures reported here are representative of a productive enzyme·NAD·D-G3P complex in the ground state (Michaelis complex).
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Contributor : Sandrine Boschi-Muller <>
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Claude Didierjean, Catherine Corbier, Mustapha Fatih, Frederique Favier, Sandrine Boschi-Muller, et al.. Crystal Structure of Two Ternary Complexes of Phosphorylating Glyceraldehyde-3-phosphate Dehydrogenase from Bacillus stearothermophilus with NAD and d-Glyceraldehyde 3-Phosphate. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2003, 278 (15), pp.12968-12976. ⟨10.1074/jbc.M211040200⟩. ⟨hal-01690824⟩

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