title: 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, Cys(149), 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 P(s) site and not in the P(i) site. Its C-1 carbonyl oxygen points toward the essential His(176), 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).
authors.affiliation: Laboratoire de Cristallographie et de Modélisation des Matériaux Minéraux et Biologiques, Groupe Biocristallographie, UMR 7036, CNRS-Université Henri Poincaré, Faculté des Sciences, 54506 Vandoeuvre Cedex, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Didierjean
authors.forename: Catherine
authors.initials: C
authors.lastname: Corbier
authors.forename: Mustapha
authors.initials: M
authors.lastname: Fatih
authors.forename: Frédérique
authors.initials: F
authors.lastname: Favier
authors.forename: Sandrine
authors.initials: S
authors.lastname: Boschi-Muller
authors.forename: Guy
authors.initials: G
authors.lastname: Branlant
authors.forename: André
authors.initials: A
authors.lastname: Aubry
mesh_terms: D019943:Amino Acid Substitution; D001665:Binding Sites; D018360:Crystallography, X-Ray; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005986:Glyceraldehyde 3-Phosphate / Q000737:chemistry* / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D005986:Glyceraldehyde 3-Phosphate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases