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J Am Chem Soc. 2015 Jan 21;137(2):556-9. doi: 10.1021/ja502109n. Epub 2015 Jan 12.

Bifunctional fatty acid chemical reporter for analyzing S-palmitoylated membrane protein-protein interactions in mammalian cells.

Author information

1
Laboratory of Chemical Biology and Bacterial Pathogenesis, The Rockefeller University , New York, New York 10065, United States.

Abstract

Studying the functions of S-palmitoylated proteins in cells can be challenging due to the membrane targeting property and dynamic nature of protein S-palmitoylation. New strategies are therefore needed to specifically capture S-palmitoylated protein complexes in cellular membranes for dissecting their functions in vivo. Here we present a bifunctional fatty acid chemical reporter, x-alk-16, which contains an alkyne and a diazirine, for metabolic labeling of S-palmitoylated proteins and photo-cross-linking of their involved protein complexes in mammalian cells. We demonstrate that x-alk-16 can be metabolically incorporated into known S-palmitoylated proteins such as H-Ras and IFITM3, a potent antiviral protein, and induce covalent cross-linking of IFITM3 oligomerization as well as its specific interactions with other membrane proteins upon in-cell photoactivation. Moreover, integration of x-alk-16-induced photo-cross-linking with label-free quantitative proteomics allows identification of new IFITM3 interacting proteins.

PMID:
25575299
PMCID:
PMC4303541
DOI:
10.1021/ja502109n
[Indexed for MEDLINE]
Free PMC Article

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