title: Further insights into the mechanism of function of the response regulator CheY from crystallographic studies of the CheY--CheA(124--257) complex.
abstract: New crystallographic structures of the response regulator CheY in association with CheA(124--257), its binding domain in the kinase CheA, have been determined. In all crystal forms, the molecular interactions at the heterodimer interface are identical. Soaking experiments have been performed on the crystals using acetyl phosphate as phosphodonor to CheY. No phosphoryl group attached to Asp57 of CheY is visible from the electron density, but the response regulator in the CheY-CheA(124--257) complex may have undergone a phosphorylation-dephosphorylation process. The distribution of water molecules and the geometry of the active site have changed and are now similar to those of isolated CheY. In a second soaking experiment, imido-diphosphate, an inhibitor of the phosphorylation reaction, was used. This compound binds in the vicinity of the active site, close to the N-terminal part of the first alpha-helix. Together, these results suggest that the binding of CheY to CheA(124--257) generates a geometry of the active site that favours phosphorylation and that imido-diphosphate interferes with phosphorylation by precluding structural changes in this region.
authors.affiliation: Groupe de Cristallographie Biologique, IPBS-CNRS, 205 Route de Narbonne, 31077 Toulouse CEDEX, France.
authors.forename: P
authors.initials: P
authors.lastname: Gouet
authors.forename: N
authors.initials: N
authors.lastname: Chinardet
authors.forename: M
authors.initials: M
authors.lastname: Welch
authors.forename: V
authors.initials: V
authors.lastname: Guillet
authors.forename: S
authors.initials: S
authors.lastname: Cabantous
authors.forename: C
authors.initials: C
authors.lastname: Birck
authors.forename: L
authors.initials: L
authors.lastname: Mourey
authors.forename: J P
authors.initials: JP
authors.lastname: Samama
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001665:Binding Sites; D018360:Crystallography, X-Ray; D008565:Membrane Proteins / Q000737:chemistry*; D000072236:Methyl-Accepting Chemotaxis Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D010766:Phosphorylation; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D010446:Peptide Fragments