Rapid and selective detection of fatty acylated proteins using ω-alkynyl-fatty acids and click chemistry [S]
MC Yap, MA Kostiuk, DDO Martin… - Journal of lipid …, 2010 - jlr.org
Progress in understanding the biology of protein fatty acylation has been impeded by the
lack of rapid direct detection and identification methods. We first report that a synthetic ω-
alkynyl-palmitate analog can be readily and specifically incorporated into GAPDH or
mitochondrial 3-hydroxyl-3-methylglutaryl-CoA synthase in vitro and reacted with an azido-
biotin probe or the fluorogenic probe 3-azido-7-hydroxycoumarin using click chemistry for
rapid detection by Western blotting or flat bed fluorescence scanning. The acylated cysteine …
lack of rapid direct detection and identification methods. We first report that a synthetic ω-
alkynyl-palmitate analog can be readily and specifically incorporated into GAPDH or
mitochondrial 3-hydroxyl-3-methylglutaryl-CoA synthase in vitro and reacted with an azido-
biotin probe or the fluorogenic probe 3-azido-7-hydroxycoumarin using click chemistry for
rapid detection by Western blotting or flat bed fluorescence scanning. The acylated cysteine …
