Alkynyl-farnesol reporters for detection of protein S-prenylation in cells.
Protein S-prenylation is a lipid modification that regulates membrane-protein and protein-protein interactions in cell signaling. Though sites of protein S-prenylation can be predicted based upon conserved C-terminal CaaX or CC/CXC motifs, biochemical detection of protein S-prenylation in cells is still challenging. Herein, we report an alkynyl-isoprenol chemical reporter (alk-FOH) as an efficient substrate for prenyltransferases in mammalian cells that enables sensitive detection of S-farnesylated and S-geranylgeranylated proteins using bioorthogonal ligation methods. Fluorescent detection alleviates the need to deplete cellular isoprenoids for biochemical analysis of S-prenylated proteins and enables robust characterization of S-prenylated proteins, such as effectors that are injected into host cells by bacterial pathogens. This alkynyl-prenylation reporter provides a sensitive tool for biochemical analysis and rapid profiling of prenylated proteins in cells.
The Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, NY 10065, USA.
Guillaume
G
Charron
Lun K
LK
Tsou
William
W
Maguire
Jacob S
JS
Yount
Howard C
HC
Hang
D000818:Animals; D002460:Cell Line; D004122:Dimethylallyltranstransferase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005204:Farnesol / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D008954:Models, Biological; D017368:Protein Prenylation / Q000502:physiology*
D005204:Farnesol; D004122:Dimethylallyltranstransferase