Abstract
SummaryCells acquire fatty acids from dietary sources or via de novo palmitate production by fatty acid synthase (FASN). Although most cells express FASN at low levels, it is upregulated in cancers and during replication of many viruses. The precise role of FASN in disease pathogenesis is poorly understood, and whether de novo fatty acid synthesis contributes to host or viral protein acylation has been traditionally difficult to study. We describe a cell permeable, click-chemistry compatible alkynyl-acetate analog (Alk-4) that functions as a reporter of FASN-dependent protein acylation. In a FASN-dependent manner, Alk-4 selectively labeled the cellular protein interferon-induced transmembrane protein 3 (IFITM3) at its palmitoylation sites, and the HIV-1 matrix protein at its myristoylation site. Alk-4 metabolic labeling also enabled biotin-based purification and identification of more than 200 FASN-dependent acylated cellular proteins. Thus, Alk-4 is a useful bioorthogonal tool to selectively probe FASN-mediated protein acylation in normal and diseased states.
Publisher
Cold Spring Harbor Laboratory
Reference59 articles.
1. Fatty acylation of proteins: The long and the short of it
2. Fatty acid metabolism: Implications for diet, genetic variation, and disease
3. Liu, H. , Liu, J. Y. , Wu, X. & Zhang, J. J. Biochemistry, molecular biology, and pharmacology of fatty acid synthase, an emerging therapeutic target and diagnosis/prognosis marker. International Journal of Biochemistry and Molecular Biology (2010).
4. Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology
5. Differential Regulation of Host Genes Including Hepatic Fatty Acid Synthase in HBV-Transgenic Mice