title: Clickable analogue of cerulenin as chemical probe to explore protein palmitoylation.
abstract: Dynamic palmitoylation is an important post-translational modification regulating protein localization, trafficking, and signaling activities. The Asp-His-His-Cys (DHHC) domain containing enzymes are evolutionarily conserved palmitoyl acyltransferases (PATs) mediating diverse protein S-palmitoylation. Cerulenin is a natural product inhibitor of fatty acid biosynthesis and protein palmitoylation, through irreversible alkylation of the cysteine residues in the enzymes. Here, we report the synthesis and characterization of a "clickable" and long alkyl chain analogue of cerulenin as a chemical probe to investigate its cellular targets and to label and profile PATs in vitro and in live cells. Our results showed that the probe could stably label the DHHC-family PATs and enable mass spectrometry studies of PATs and other target proteins in the cellular proteome. Such probe provides a new chemical tool to dissect the functions of palmitoylating enzymes in cell signaling and diseases and reveals new cellular targets of the natural product cerulenin.
authors.affiliation: Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School , Building 149, 13th Street, Charlestown, Massachusetts 02129, United States.
authors.forename: Baohui
authors.initials: B
authors.lastname: Zheng
authors.forename: Shunying
authors.initials: S
authors.lastname: Zhu
authors.forename: Xu
authors.initials: X
authors.lastname: Wu
mesh_terms: D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000737:chemistry / Q000378:metabolism; D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002569:Cerulenin / Q000031:analogs & derivatives / Q000138:chemical synthesis; D057930:Click Chemistry; D006801:Humans; D054878:Lipoylation; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013869:Thiolester Hydrolases / Q000737:chemistry / Q000378:metabolism
chemical_list: D015335:Molecular Probes; D011506:Proteins; D002569:Cerulenin; D000217:Acyltransferases; D008243:1-Acylglycerophosphocholine O-Acyltransferase; C586147:LPCAT2 protein, human; C402226:LYPLA1 protein, human; D013869:Thiolester Hydrolases