title: Carbamoyl phosphate synthetase: a crooked path from substrates to products.
abstract: The formation of carbamoyl phosphate is catalyzed by a single enzyme using glutamine, bicarbonate and two molecules of ATP via a reaction mechanism that requires a minimum of four consecutive reactions and three unstable intermediates. The recently determined X-ray crystal structure of carbamoyl phosphate synthetase has revealed the location of three separate active sites connected by two molecular tunnels that run through the interior of the protein. It has been demonstrated that the amidotransferase domain within the small subunit of the enzyme from Escherichia coli hydrolyzes glutamine to ammonia via a thioester intermediate with Cys269. The ammonia migrates through the interior of the protein, where it reacts with carboxy phosphate to produce the carbamate intermediate. The carboxy phosphate intermediate is formed by the phosphorylation of bicarbonate by ATP at a site contained within the amino-terminal half of the large subunit. The carbamate intermediate is transported through the interior of the protein to a second site within the carboxy-terminal half of the large subunit, where it is phosphorylated by another ATP to yield the final product, carbamoyl phosphate. The entire journey from substrate to product covers a distance of nearly 100 A.
authors.affiliation: Department of Chemistry, Texas A&M University, College Station 77843, USA. raushel@tamu.edu
authors.forename: F M
authors.initials: FM
authors.lastname: Raushel
authors.forename: J B
authors.initials: JB
authors.lastname: Thoden
authors.forename: G D
authors.initials: GD
authors.lastname: Reinhart
authors.forename: H M
authors.initials: HM
authors.lastname: Holden
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000494:Allosteric Regulation / Q000502:physiology; D000495:Allosteric Site / Q000502:physiology; D000595:Amino Acid Sequence; D001639:Bicarbonates / Q000378:metabolism; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / Q000737:chemistry*; D002221:Carbamyl Phosphate / Q000737:chemistry*; D020134:Catalytic Domain / Q000502:physiology; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D005973:Glutamine / Q000378:metabolism; D007527:Isoenzymes; D012045:Regulatory Sequences, Nucleic Acid
chemical_list: D001639:Bicarbonates; D007527:Isoenzymes; D005973:Glutamine; D002221:Carbamyl Phosphate; D000255:Adenosine Triphosphate; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)