1kee

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1kee, resolution 2.10Å ()
Ligands: , , , , , ,
Non-Standard Residues:
Gene: CARB (Escherichia coli), CARA (Escherichia coli)
Activity: Carbamoyl-phosphate synthase (glutamine-hydrolyzing), with EC number 6.3.5.5
Resources: FirstGlance, OCA, RCSB, PDBsum
Coordinates: save as pdb, mmCIF, xml



Contents

Inactivation of the Amidotransferase Activity of Carbamoyl Phosphate Synthetase by the Antibiotic Acivicin

Publication Abstract from PubMed

Carbamoyl phosphate synthetase (CPS) from Escherichia coli catalyzes the formation of carbamoyl phosphate from 2 mol of ATP, bicarbonate, and glutamine. CPS was inactivated by the glutamine analog, acivicin. In the presence of ATP and bicarbonate the second-order rate constant for the inactivation of the glutamine-dependent activities was 4.0 x 10(4) m(-1) s(-1). In the absence of ATP and bicarbonate the second-order rate constant for inactivation of CPS was reduced by a factor of 200. The enzyme was protected against inactivation by the inclusion of glutamine in the reaction mixture. The ammonia-dependent activities were unaffected by the incubation of CPS with acivicin. These results are consistent with the covalent labeling of the glutamine-binding site located within the small amidotransferase subunit. The binding of ATP and bicarbonate to the large subunit of CPS must also induce a conformational change within the amidotransferase domain of the small subunit that enhances the nucleophilic character of the thiol group required for glutamine hydrolysis. The acivicin-inhibited enzyme was crystallized, and the three-dimensional structure was determined by x-ray diffraction techniques. The thiol group of Cys-269 was covalently attached to the dihydroisoxazole ring of acivicin with the displacement of a chloride ion.

Inactivation of the amidotransferase activity of carbamoyl phosphate synthetase by the antibiotic acivicin., Miles BW, Thoden JB, Holden HM, Raushel FM, J Biol Chem. 2002 Feb 8;277(6):4368-73. Epub 2001 Nov 29. PMID:11729189

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

About this Structure

1kee is a 8 chain structure with sequence from Escherichia coli. Full crystallographic information is available from OCA.

See Also

Reference

  • Miles BW, Thoden JB, Holden HM, Raushel FM. Inactivation of the amidotransferase activity of carbamoyl phosphate synthetase by the antibiotic acivicin. J Biol Chem. 2002 Feb 8;277(6):4368-73. Epub 2001 Nov 29. PMID:11729189 doi:10.1074/jbc.M108582200

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