Source:http://linkedlifedata.com/resource/pubmed/id/15491158
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
42
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| pubmed:dateCreated |
2004-10-19
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| pubmed:abstractText |
Cyclopropane fatty acid (CFA) synthases catalyze the formation of cyclopropane rings on unsaturated fatty acids (UFAs) that are natural components of membrane phospholipids. The methylene carbon of the cyclopropane ring derives from the activated methyl group of S-adenosyl-L-methionine (AdoMet), affording S-adenosyl-L-homocysteine (AdoHcys) and a proton as the remaining products. This reaction is unique among AdoMet-dependent enzymes, because the olefin of the UFA substrate is isolated and unactivated toward nucleophilic or electrophilic addition, raising the question as to the timing and mechanism of proton loss from the activated methyl group of AdoMet. Two distinct reaction schemes have been proposed for this transformation; however, neither was based on detailed in vitro mechanistic analysis of the enzyme. In the preceding paper [Iwig, D. F. and Booker, S. J. (2004) Biochemistry 43, http://dx.doi.org/10.1021/bi048693+], we described the synthesis of two analogues of AdoMet, Se-adenosyl-L-selenomethionine (SeAdoMet) and Te-adenosyl-L-telluromethionine (TeAdoMet), and their intrinsic reactivity toward polar chemistry in which AdoMet is known to be involved. We found that the electrophilicity of AdoMet and its onium congeners followed the series SeAdoMet > AdoMet > TeAdoMet, while the acidity of the carbons adjacent to the relevant heteroatom followed the series AdoMet > SeAdoMet > TeAdoMet. When each of these compounds was used as the methylene donor in the CFA synthase reaction, the kinetic parameters of the reaction, k(cat) and k(cat) K(M)(-1), followed the series SeAdoMet > AdoMet > TeAdoMet, suggesting that the reaction takes place via methyl transfer followed by proton loss, rather than by processes that are initiated by proton abstraction from AdoMet. Use of S-adenosyl-L-[methyl-d(3)]methionine as the methylene donor resulted in an inverse isotope effect of 0.87 +/- 0.083, supporting this conclusion and also indicating that the methyl transfer takes place via a tight s(N)2 transition state.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Catechol O-Methyltransferase,
http://linkedlifedata.com/resource/pubmed/chemical/Chelating Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Cyclopropanes,
http://linkedlifedata.com/resource/pubmed/chemical/Escherichia coli Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Fatty Acids,
http://linkedlifedata.com/resource/pubmed/chemical/Metals, Heavy,
http://linkedlifedata.com/resource/pubmed/chemical/Methionine,
http://linkedlifedata.com/resource/pubmed/chemical/Methyltransferases,
http://linkedlifedata.com/resource/pubmed/chemical/Phospholipids,
http://linkedlifedata.com/resource/pubmed/chemical/Protons,
http://linkedlifedata.com/resource/pubmed/chemical/S-Adenosylmethionine,
http://linkedlifedata.com/resource/pubmed/chemical/Selenomethionine,
http://linkedlifedata.com/resource/pubmed/chemical/Tellurium,
http://linkedlifedata.com/resource/pubmed/chemical/adenosylselenomethionine,
http://linkedlifedata.com/resource/pubmed/chemical/cyclopropane fatty acids,
http://linkedlifedata.com/resource/pubmed/chemical/cyclopropane synthetase,
http://linkedlifedata.com/resource/pubmed/chemical/telluromethionine
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| pubmed:status |
MEDLINE
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| pubmed:month |
Oct
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| pubmed:issn |
0006-2960
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
26
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| pubmed:volume |
43
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
13510-24
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:15491158-Catalysis,
pubmed-meshheading:15491158-Catechol O-Methyltransferase,
pubmed-meshheading:15491158-Chelating Agents,
pubmed-meshheading:15491158-Cyclopropanes,
pubmed-meshheading:15491158-Deuterium Exchange Measurement,
pubmed-meshheading:15491158-Escherichia coli Proteins,
pubmed-meshheading:15491158-Fatty Acids,
pubmed-meshheading:15491158-Kinetics,
pubmed-meshheading:15491158-Metals, Heavy,
pubmed-meshheading:15491158-Methionine,
pubmed-meshheading:15491158-Methyltransferases,
pubmed-meshheading:15491158-Models, Chemical,
pubmed-meshheading:15491158-Phospholipids,
pubmed-meshheading:15491158-Protons,
pubmed-meshheading:15491158-S-Adenosylmethionine,
pubmed-meshheading:15491158-Selenomethionine,
pubmed-meshheading:15491158-Spectrophotometry, Ultraviolet,
pubmed-meshheading:15491158-Substrate Specificity,
pubmed-meshheading:15491158-Tellurium
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| pubmed:year |
2004
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| pubmed:articleTitle |
Isotope and elemental effects indicate a rate-limiting methyl transfer as the initial step in the reaction catalyzed by Escherichia coli cyclopropane fatty acid synthase.
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| pubmed:affiliation |
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
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| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.
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