| Analysis of protein S-acylation by gas chromatography-coupled mass spectrometry using purified proteins. | |
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MedLine Citation:
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PMID: 20379138 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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S-acylation, also known as palmitoylation, involves the attachment of acyl fatty acids to thiol groups of cysteine residues through a reversible thioester bond. Owing to its reversibility, S-acylation is important in regulation of diverse signaling cascades, including Ras-associated cancers in mammals, stress response and metabolic regulation. Here we describe a simple protocol for analysis of protein S-acylation using gas chromatography-coupled mass spectrometry. Analysis can be carried out with as little as 1 microg of purified protein and allows chemical identification and, potentially, quantification of the acyl moieties. The method is based on cleavage of the fatty acids from proteins by hydrogenation with platinum (IV) oxide. This causes an acid transesterification of the acyl groups, adding an ethyl group to the carboxyl head of the fatty acid. The addition of the ethyl group reduces the polarity of the fatty acids, allowing their efficient separation by gas chromatography. |
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Authors:
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Nadav Sorek; Shaul Yalovsky |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-04-08 |
Journal Detail:
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Title: Nature protocols Volume: 5 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2010 May |
Date Detail:
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Created Date: 2010-04-09 Completed Date: 2010-06-29 Revised Date: 2012-03-30 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 834-40 Citation Subset: IM |
Affiliation:
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Department of Plant Sciences, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acylation Esterification Gas Chromatography-Mass Spectrometry / methods* Lipoylation Proteins / chemistry*, isolation & purification* Proteomics / methods |
| Chemical | |
Reg. No./Substance:
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0/Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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