title: New insights into the mechanism of nucleoside hydrolases from the crystal structure of the Escherichia coli YbeK protein bound to the reaction product.
abstract: Nucleoside hydrolases (NHs) are enzymes that catalyze the excision of the N-glycosidic bond in nucleosides to allow recycling of the nitrogenous bases. The fine details of the catalytic mechanism and the structural features imposing the substrate specificity of the various members of the NH family are still debated. Here we present the functional characterization of the Escherichia coli YbeK (RihA) protein as a pyrimidine nucleoside-preferring NH and its first crystal structure to 1.8 A resolution. The enzyme active site is occupied by either the alpha or beta anomer of ribose and provides the first structural description of the binding of the NH reaction product. While the amino acid residues involved in ribosyl binding are strictly conserved in pyrimidine-specific NHs, the residues involved in specific interactions with the nitrogenous bases differ considerably. Further comparison of the active site architecture of YbeK with the related NHs establishes structural determinants involved in triggering the conformational transition between the open and closed structures and suggests a mechanism for product release.
authors.affiliation: Biocrystallography Unit, DIBIT Scientific Institute S. Raffaele, via Olgettina 58, 20132 Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Muzzolini
authors.forename: Wim
authors.initials: W
authors.lastname: Versées
authors.forename: Paola
authors.initials: P
authors.lastname: Tornaghi
authors.forename: Els
authors.initials: E
authors.lastname: Van Holsbeke
authors.forename: Jan
authors.initials: J
authors.lastname: Steyaert
authors.forename: Massimo
authors.initials: M
authors.lastname: Degano
mesh_terms: D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D009699:N-Glycosyl Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012266:Ribose / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D012266:Ribose; D009699:N-Glycosyl Hydrolases; C422987:RihA protein, E coli