The development of tools for proteomic analysis is an active area of research. Here, we report on the synthesis of 12-propargoxyfarnesyl diphosphate (1), an alkyne-containing analogue of farnesyl diphosphate (FPP), and its enzymatic incorporation into peptide substrates by both protein- farnesyltransferase (PFTase) and protein-geranylgeranyltransferase type I (PGGTase-I). Compound 1 was prepared from farnesol in 6 steps. Kinetic analyses indicate that 1 is incorporated into cognate peptide substrates by PFTase or PGGTase at concentrations and rates comparable to those of the natural lipid substrates for these enzymes, and mass spectrometric analyses proved the structures of the prenylated peptide products. Incubation of 1 in the presence of PFTase and PGGTase peptide substrates, and the cognate transferases, results in the simultaneous prenylation of both peptides emphasizing the dual substrate nature of 1. Thus, because 1 is a substrate for both enzymes, it can be used to introduce alkyne functionality into proteins that are normally either farnesylated or geranylgeranylated. This approach should be useful for a broad range of applications ranging from selective protein labeling to proteomic analysis.
|Original language||English (US)|
|Number of pages||10|
|Journal||International Journal of Peptide Research and Therapeutics|
|State||Published - Jun 2007|
Bibliographical noteFunding Information:
The authors thank Dr. Tom Krick, Dr. Lee Ann Higgins, and Sean Murray in the Biological Mass Spectrometry Laboratory at the University of Minnesota for their assistance in obtaining the ESI-MS data reported here, and Dr. Daniel G. Mullen for his help with peptide synthesis. This work was supported by a grant from the National Institutes of Health (GM58442).
- Peptide modification
- Prenylated peptide