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| - | [[Image:1cla.jpg|left|200px]] | + | {{Seed}} |
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| | {{STRUCTURE_1cla| PDB=1cla | SCENE= }} | | {{STRUCTURE_1cla| PDB=1cla | SCENE= }} |
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| - | '''EVIDENCE FOR TRANSITION-STATE STABILIZATION BY SERINE-148 IN THE CATALYTIC MECHANISM OF CHLORAMPHENICOL ACETYLTRANSFERASE'''
| + | ===EVIDENCE FOR TRANSITION-STATE STABILIZATION BY SERINE-148 IN THE CATALYTIC MECHANISM OF CHLORAMPHENICOL ACETYLTRANSFERASE=== |
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| - | ==Overview==
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| - | The function of conserved Ser-148 of chloramphenicol acetyltransferase (CAT) has been investigated by site-directed mutagenesis. Modeling studies (P. C. E. Moody and A. G. W. Leslie, unpublished results) suggested that the hydroxyl group of Ser-148 could be involved in transition-state stabilization via a hydrogen bond to the oxyanion of the putative tetrahedral intermediate. Replacement of serine by alanine results in a mutant enzyme (Ala-148 CAT) with kcat reduced 53-fold and only minor changes in Km values for chloramphenicol and acetyl-CoA. The Ser-148----Gly substitution gives rise to a mutant enzyme (Gly-148 CAT) with kcat reduced only 10-fold. A water molecule may partially replace the hydrogen-bonding potential of Ser-148 in Gly-148 CAT. The three-dimensional structure of Ala-148 CAT at 2.34-A resolution is isosteric with that of wild-type CAT with two exceptions: the absence of the Ser-148 hydroxyl group and the loss of one poorly ordered water molecule from the active site region. The results are consistent with a catalytic role for Ser-148 rather than a structural one and support the hypothesis that Ser-148 is involved in transition-state stabilization. Ser-148 has also been replaced with cysteine and asparagine; the Ser-148----Cys mutation results in a 705-fold decrease in kcat and the Ser-148----Asn substitution in a 214-fold reduction in kcat. Removing the hydrogen bond donor (Ser-148----Ala or Gly) is less deleterious than replacing Ser-148 with alternative possible hydrogen bond donors (Ser-148----Cys or Asn). | + | The line below this paragraph, {{ABSTRACT_PUBMED_2109633}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 2109633 is the PubMed ID number. |
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| | + | {{ABSTRACT_PUBMED_2109633}} |
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| | ==About this Structure== | | ==About this Structure== |
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| | [[Category: Gibbs, M R.]] | | [[Category: Gibbs, M R.]] |
| | [[Category: Leslie, A G.W.]] | | [[Category: Leslie, A G.W.]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 12:51:30 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jun 30 20:54:25 2008'' |
Revision as of 17:54, 30 June 2008
Template:STRUCTURE 1cla
EVIDENCE FOR TRANSITION-STATE STABILIZATION BY SERINE-148 IN THE CATALYTIC MECHANISM OF CHLORAMPHENICOL ACETYLTRANSFERASE
Template:ABSTRACT PUBMED 2109633
About this Structure
1CLA is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
Reference
Evidence for transition-state stabilization by serine-148 in the catalytic mechanism of chloramphenicol acetyltransferase., Lewendon A, Murray IA, Shaw WV, Gibbs MR, Leslie AG, Biochemistry. 1990 Feb 27;29(8):2075-80. PMID:2109633
Page seeded by OCA on Mon Jun 30 20:54:25 2008
