title: Bioorthogonal proteomics of 15-hexadecynyloxyacetic acid chemical reporter reveals preferential targeting of fatty acid modified proteins and biosynthetic enzymes.
abstract: Chemical reporters are powerful tools for the detection and discovery of protein modifications following cellular labeling. The metabolism of alkyne- or azide-functionalized chemical reporters in cells can influence the efficiency and specificity of protein targeting. To evaluate the effect of degradation of chemical reporters of protein fatty acylation, we synthesized 15-hexadecynyloxyacetic acid (HDYOA), a reporter that was designed to be resistant to β-oxidation, and compared its ability to label palmitoylated proteins with an established reporter, 17-octadecynoic acid (ODYA). HDYOA was able to label known candidate S-palmitoylated proteins similarly to ODYA. Accordingly, bioorthogonal proteomic analysis demonstrated that 70% of proteins labeled with ODYA were also labeled with HDYOA. However, the proteins observed differentially in our proteomic studies suggested that a portion of ODYA protein labeling is a result of β-oxidation. In contrast, downstream enzymes involved in β-oxidation of fatty acids were not targeted by HDYOA. Since HDYOA can label S-palmitoylated proteins and is not utilized by downstream β-oxidation pathways, this fatty acid chemical reporter may be particularly useful for bioorthogonal proteomic studies in cell types metabolically skewed toward fatty acid breakdown.
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, 1230 York Avenue, Box 250, New York, NY 10065, United States.
authors.forename: Jacob S
authors.initials: JS
authors.lastname: Yount
authors.forename: Guillaume
authors.initials: G
authors.lastname: Charron
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
mesh_terms: D000085:Acetates / Q000138:chemical synthesis / Q000737:chemistry*; D000217:Acyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D010084:Oxidation-Reduction; D040901:Proteomics
chemical_list: C572003:15-hexadecynyloxyacetic acid; D000085:Acetates; D005231:Fatty Acids, Unsaturated; C047008:17-octadecynoic acid; D000217:Acyltransferases; C074049:fatty acid modifying enzyme