title: Bifunctional fatty acid chemical reporter for analyzing S-palmitoylated membrane protein-protein interactions in mammalian cells.
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.
authors.affiliation: Laboratory of Chemical Biology and Bacterial Pathogenesis, The Rockefeller University , New York, New York 10065, United States.
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
mesh_terms: D000480:Alkynes / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D003978:Diazomethane / Q000737:chemistry; D005227:Fatty Acids / Q000737:chemistry* / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D054878:Lipoylation; D008565:Membrane Proteins / Q000378:metabolism*; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D000480:Alkynes; D005227:Fatty Acids; D008565:Membrane Proteins; D015335:Molecular Probes; D003978:Diazomethane