title: Structural basis for the feedback regulation of Escherichia coli pantothenate kinase by coenzyme A.
abstract: Pantothenate kinase (PanK) is a key regulatory enzyme in the coenzyme A (CoA) biosynthetic pathway and catalyzes the phosphorylation of pantothenic acid to form phosphopantothenate. CoA is a feedback inhibitor of PanK activity by competitive binding to the ATP site. The structures of the Escherichia coli enzyme, in complex with a nonhydrolyzable analogue of ATP, 5'-adenylimido-diphosphate (AMPPNP), or with CoA, were determined at 2.6 and 2.5 A, respectively. Both structures show that two dimers occupy an asymmetric unit; each subunit has a alpha/beta mononucleotide-binding fold with an extensive antiparallel coiled coil formed by two long helices along the dimerization interface. The two ligands, AMPPNP and CoA, associate with PanK in very different ways, but their phosphate binding sites overlap, explaining the kinetic competition between CoA and ATP. Residues Asp(127), His(177), and Arg(243) are proposed to be involved in catalysis, based on modeling of the pentacoordinate transition state. The more potent inhibition by CoA, compared with the CoA thioesters, is explained by a tight interaction of the CoA thiol group with the side chains of aromatic residues, which is predicted to discriminate against the CoA thioesters. The PanK structure provides the framework for a more detailed understanding of the mechanism of catalysis and feedback regulation of PanK.
authors.affiliation: Departments of Structural Biology and Biochemistry, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors.forename: M
authors.initials: M
authors.lastname: Yun
authors.forename: C G
authors.initials: CG
authors.lastname: Park
authors.forename: J Y
authors.initials: JY
authors.lastname: Kim
authors.forename: C O
authors.initials: CO
authors.lastname: Rock
authors.forename: S
authors.initials: S
authors.lastname: Jackowski
authors.forename: H W
authors.initials: HW
authors.lastname: Park
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology*; D000595:Amino Acid Sequence; D001120:Arginine; D001224:Aspartic Acid; D001665:Binding Sites; D003065:Coenzyme A / Q000378:metabolism*; D018360:Crystallography, X-Ray; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology*; D005246:Feedback; D006639:Histidine; D008024:Ligands; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary
chemical_list: D008024:Ligands; D000266:Adenylyl Imidodiphosphate; D001224:Aspartic Acid; D006639:Histidine; D000255:Adenosine Triphosphate; D001120:Arginine; D017853:Phosphotransferases (Alcohol Group Acceptor); C023088:pantothenate kinase; D003065:Coenzyme A