X-ray crystal structure of aminoimidazole ribonucleotide synthetase (PurM), from the Escherichia coli purine biosynthetic pathway at 2.5 angstrom resolution

Background: The purine biosynthetic pathway in procaryotes enlists eleven e nzymes, six of which use ATP. Enzymes 5 and 6 of this pathway, formylglycin amide ribonucleotide (FGAR) amidotransferase (Purl) and aminoimidazole ribo nucleotide (AIR) synthetase (PurM) utilize ATP to activate the oxygen of an amide within their substrate toward nucleophilic attack by a nitrogen. AIR synthetase uses the product of Purl, formylglycinamidine ribonucleotide (F GAM) and ATP to make AIR, ADP and P-j. Results: The structure of a hexahistidine-tagged PurM has been solved by mu ltiwavelength anomalous diffraction phasing techniques using protein contai ning 28 selenomethionines per asymmetric unit. The final model of PurM cons ists of two crystallographically independent dimers and four sulfates. The overall R factor at 2.5 Angstrom resolution is 19.2%, with an R-free of 26. 4%. The active site, identified in part by conserved residues, is proposed to be a long groove generated by the interaction of two monomers, A search of the sequence databases suggests that the ATP-binding sites between PurM and Purl may be structurally conserved. Conclusions: The first structure of a new class of ATP-binding enzyme, PurM , has been solved and a model for the active site has been proposed. The st ructure is unprecedented, with an extensive and unusual sheet-mediated inte rsubunit interaction defining the active-site grooves. Sequence searches su ggest that two successive enzymes in the purine biosynthetic pathway, propo sed to use similar chemistries, will have similar ATP-binding domains.