research papers Acta Crystallographica Section D Biological The structure of carbamoyl phosphate synthetase Crystallography determined to 2.1 AÊ resolution ISSN 0907-4449 James B. Thoden,a Frank M. Carbamoyl phosphate synthetase catalyzes the formation of Received 10 March 1998 Raushel,b Matthew M. Benning,a carbamoyl phosphate from one molecule of bicarbonate, two Accepted 29 April 1998 2+ Ivan Raymenta and Hazel M. molecules of Mg ATP and one molecule of glutamine or a ammonia depending upon the particular form of the enzyme PDB Reference: carbamoyl Holden * phosphate synthetase, 1jdb. under investigation. As isolated from Escherichia coli, the enzyme is an , -heterodimer consisting of a small subunit aInstitute for Enzyme Research, The Graduate that hydrolyzes glutamine and a large subunit that catalyzes School, and Department of Biochemistry, the two required phosphorylation events. Here the three- College of Agricultural and Life Sciences, University of Wisconsin±Madison, 1710 dimensional structure of carbamoyl phosphate synthetase University Avenue, Madison, Wisconsin 53705, from E. coli re®ned to 2.1 AÊ resolution with an R factor of USA, and bDepartment of Chemistry, Texas 17.9% is described. The small subunit is distinctly bilobal with A&M University, College Station, Texas 77843, a catalytic triad (Cys269, His353 and Glu355) situated USA between the two structural domains. As observed in those enzymes belonging to the = -hydrolase family, the active-site Correspondence e-mail: nucleophile, Cys269, is perched at the top of a tight turn. The
[email protected] large subunit consists of four structural units: the carboxypho- sphate synthetic component, the oligomerization domain, the carbamoyl phosphate synthetic component and the allosteric domain.