The Activity of Escherichia coli Cyclopropane Fatty Acid Synthase Depends on the Presence of Bicarbonate

Cyclopropane fatty acid (CFA) synthases catalyze the formation of cyclopropane rings on isolated and unactivated olefinic bonds within various fatty acids; the methylene carbon is derived from the activated methyl group of (S)-adenosylmethionine. The E. coli enzyme is the prototype for this class of...

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Veröffentlicht in:Journal of the American Chemical Society 2005-08, Vol.127 (33), p.11612-11613
Hauptverfasser: Iwig, David F, Uchida, Akira, Stromberg, Jeffrey A, Booker, Squire J
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container_end_page 11613
container_issue 33
container_start_page 11612
container_title Journal of the American Chemical Society
container_volume 127
creator Iwig, David F
Uchida, Akira
Stromberg, Jeffrey A
Booker, Squire J
description Cyclopropane fatty acid (CFA) synthases catalyze the formation of cyclopropane rings on isolated and unactivated olefinic bonds within various fatty acids; the methylene carbon is derived from the activated methyl group of (S)-adenosylmethionine. The E. coli enzyme is the prototype for this class of enzymes, which include the cyclopropane mycolic acid (CMA) synthases, which are potential targets for the design of antituberculosis agents. Crystal structures of several CMA synthases have recently been solved, and electron density attributed to a bicarbonate ion was found in or near the active site. Because a functional assay for CMA synthases has not been developed, the relevance of the bicarbonate ion has not been established. CFA synthase is 30−35% identical to the CMA synthases that have been analyzed structurally, suggesting that the mechanisms of these enzymes are conserved. In this work, we show that indeed the activity of CFA synthase requires bicarbonate, and that it is inhibited by borate, a planar trigonal molecule that mimics the structure of bicarbonate. We also show that substitutions of the conserved amino acids that act as ligands to the bicarbonate ion based on the structure of CMA synthases result in drastic losses in the activity of the protein.
doi_str_mv 10.1021/ja053899z
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subjects Bicarbonates - chemistry
Bicarbonates - pharmacology
Biological and medical sciences
Escherichia coli - enzymology
Fundamental and applied biological sciences. Psychology
Mechanisms. Catalysis. Electron transfer. Models
Methyltransferases - chemistry
Methyltransferases - drug effects
Methyltransferases - metabolism
Molecular biophysics
Molecular Structure
Physical chemistry in biology
title The Activity of Escherichia coli Cyclopropane Fatty Acid Synthase Depends on the Presence of Bicarbonate
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