title: IMP Alone Organizes the Active Site of Adenylosuccinate Synthetase from Escherichia coli.
abstract: A complete set of substrate/substrate analogs of adenylosuccinate synthetase from Escherichia coli induces dimer formation and a transition from a disordered to an ordered active site. The most striking of the ligand-induced effects is the movement of loop 40-53 by up to 9 A. Crystal structures of the partially ligated synthetase, which either combine IMP and hadacidin or IMP, hadacidin, and Mg(2+)-pyrophosphate, have ordered active sites, comparable with the fully ligated enzyme. More significantly, a crystal structure of the synthetase with IMP alone exhibits a largely ordered active site, which includes the 9 A movement of loop 40-53 but does not include conformational adjustments to backbone carbonyl 40 (Mg(2+) interaction element) and loop 298-304 (L-aspartate binding element). Interactions involving the 5'-phosphoryl group of IMP evidently trigger the formation of salt links some 30 A away. The above provides a structural basis for ligand binding synergism, effects on k(cat) due to mutations far from the site of catalysis, and the complete loss of substrate efficacy due to minor alterations of the 5'-phosphoryl group of IMP.
authors.affiliation: Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: Zhenglin
authors.initials: Z
authors.lastname: Hou
authors.forename: Wenyan
authors.initials: W
authors.lastname: Wang
authors.forename: Herbert J
authors.initials: HJ
authors.lastname: Fromm
authors.forename: Richard B
authors.initials: RB
authors.lastname: Honzatko
mesh_terms: D000265:Adenylosuccinate Synthase / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000378:metabolism; D007291:Inosine Monophosphate / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation
chemical_list: D007291:Inosine Monophosphate; D006160:Guanosine Triphosphate; C001321:hadacidin; D000265:Adenylosuccinate Synthase; D005998:Glycine