1gce: Difference between revisions
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==Overview== | ==Overview== | ||
A class C beta-lactamase from a clinical isolate of Enterobacter cloacae, strain GC1 with improved hydrolytic activity for oxyimino beta-lactam, antibiotics has been analyzed by X-ray crystallography to 1.8 A, resolution. Relative to the wild-type P99 beta-lactamase, this natural, mutant contains a highly unique tandem repeat Ala211-Val212-Arg213 [Nugaka, et al. (1995) J. Biol. Chem. 270, 5729-5735]. The 39.4 kDa chromosomal, beta-lactamase crystallizes from poly(ethylene glycol) 8000 in potassium, phosphate in space group P2(1)2(1)2 with cell dimensions a = 78.0 A, b =, 69.5 A, and c = 63.1 A. The crystal structure was solved by the molecular, replacement method, and the model has been refined to an R-factor of 0.20, for all nonzero data from 8 to 1.8 A. Deviations of model bonds and ... | A class C beta-lactamase from a clinical isolate of Enterobacter cloacae, strain GC1 with improved hydrolytic activity for oxyimino beta-lactam, antibiotics has been analyzed by X-ray crystallography to 1.8 A, resolution. Relative to the wild-type P99 beta-lactamase, this natural, mutant contains a highly unique tandem repeat Ala211-Val212-Arg213 [Nugaka, et al. (1995) J. Biol. Chem. 270, 5729-5735]. The 39.4 kDa chromosomal, beta-lactamase crystallizes from poly(ethylene glycol) 8000 in potassium, phosphate in space group P2(1)2(1)2 with cell dimensions a = 78.0 A, b =, 69.5 A, and c = 63.1 A. The crystal structure was solved by the molecular, replacement method, and the model has been refined to an R-factor of 0.20, for all nonzero data from 8 to 1.8 A. Deviations of model bonds and angles, from ideal values are 0.008 A and 1.4 degrees, respectively. Overlay of, alpha-carbon atoms in the GC1 and P99 beta-lactamases results in an rms, deviation of 0.6 A. Largest deviations occur in a loop containing Gln120, and in the Omega loop region (200-218) where the three residues 213-215, are disordered. Possibly as a result of this disorder, the width of the, opening to the substrate binding cavity, as measured from the 318-324, beta-strand to two loops containing Gln120 and Tyr150 on the other side, is 0.6-1.4 A wider than in P99. It is suggested that conformational, flexibility in the expanded Omega loop, and its influence on adjacent, protein structure, may facilitate hydrolysis of oxyimino beta-lactams by, making the acyl intermediate more open to attack by water. Nevertheless, backbone atoms in core catalytic site residues Ser64, Lys67, Tyr150, Asn152, Lys318, and Ser321 deviate only 0.4 A (rmsd) from atoms in P99. A, rotation of a potential catalytic base, Tyr150, relative to P99 at pH 8, is consistent with the requirement for a lower than normal pK(a) for this, residue. | ||
==About this Structure== | ==About this Structure== | ||
1GCE is a | 1GCE is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Enterobacter_cloacae Enterobacter cloacae]. Active as [http://en.wikipedia.org/wiki/Beta-lactamase Beta-lactamase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.6 3.5.2.6] Structure known Active Site: ASA. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1GCE OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: extended-spectrum beta- lactamase]] | [[Category: extended-spectrum beta- lactamase]] | ||
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Revision as of 16:20, 5 November 2007
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STRUCTURE OF THE BETA-LACTAMASE OF ENTEROBACTER CLOACAE GC1
OverviewOverview
A class C beta-lactamase from a clinical isolate of Enterobacter cloacae, strain GC1 with improved hydrolytic activity for oxyimino beta-lactam, antibiotics has been analyzed by X-ray crystallography to 1.8 A, resolution. Relative to the wild-type P99 beta-lactamase, this natural, mutant contains a highly unique tandem repeat Ala211-Val212-Arg213 [Nugaka, et al. (1995) J. Biol. Chem. 270, 5729-5735]. The 39.4 kDa chromosomal, beta-lactamase crystallizes from poly(ethylene glycol) 8000 in potassium, phosphate in space group P2(1)2(1)2 with cell dimensions a = 78.0 A, b =, 69.5 A, and c = 63.1 A. The crystal structure was solved by the molecular, replacement method, and the model has been refined to an R-factor of 0.20, for all nonzero data from 8 to 1.8 A. Deviations of model bonds and angles, from ideal values are 0.008 A and 1.4 degrees, respectively. Overlay of, alpha-carbon atoms in the GC1 and P99 beta-lactamases results in an rms, deviation of 0.6 A. Largest deviations occur in a loop containing Gln120, and in the Omega loop region (200-218) where the three residues 213-215, are disordered. Possibly as a result of this disorder, the width of the, opening to the substrate binding cavity, as measured from the 318-324, beta-strand to two loops containing Gln120 and Tyr150 on the other side, is 0.6-1.4 A wider than in P99. It is suggested that conformational, flexibility in the expanded Omega loop, and its influence on adjacent, protein structure, may facilitate hydrolysis of oxyimino beta-lactams by, making the acyl intermediate more open to attack by water. Nevertheless, backbone atoms in core catalytic site residues Ser64, Lys67, Tyr150, Asn152, Lys318, and Ser321 deviate only 0.4 A (rmsd) from atoms in P99. A, rotation of a potential catalytic base, Tyr150, relative to P99 at pH 8, is consistent with the requirement for a lower than normal pK(a) for this, residue.
About this StructureAbout this Structure
1GCE is a Single protein structure of sequence from Enterobacter cloacae. Active as Beta-lactamase, with EC number 3.5.2.6 Structure known Active Site: ASA. Full crystallographic information is available from OCA.
ReferenceReference
Structure of the extended-spectrum class C beta-lactamase of Enterobacter cloacae GC1, a natural mutant with a tandem tripeptide insertion., Crichlow GV, Kuzin AP, Nukaga M, Mayama K, Sawai T, Knox JR, Biochemistry. 1999 Aug 10;38(32):10256-61. PMID:10441119
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