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Quantitative Determination of Cellular Farnesyltransferase Activity: Towards Defining the Minimum Substrate Reactivity for Biologically Relevant Protein Farnesylation
Author(s) -
Flynn Susan C.,
Lindgren Danielle E.,
Hougland James L.
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402239
Subject(s) - farnesyltransferase , prenylation , prenyltransferase , geranylgeranylation , in vivo , biochemistry , in vitro , proteome , farnesyltransferase inhibitor , chemistry , biology , enzyme , microbiology and biotechnology
Abstract Prenylation is a post‐translational modification wherein an isoprenoid group is attached to a protein substrate by a protein prenyltransferase. Hundreds of peptide sequences are in vitro substrates for protein farnesyltransferase (FTase), but it remains unknown which of these sequences can successfully compete for in vivo prenylation. Translating in vitro studies to predict in vivo protein farnesylation requires determining the minimum reactivity needed for modification by FTase within the cell. Towards this goal, we developed a reporter protein series spanning several orders of magnitude in FTase reactivity as a calibrated sensor for endogenous FTase activity. Our approach provides a minimally invasive method to monitor changes in cellular FTase activity in response to environmental or genetic factors. Determining the reactivity “threshold” for in vivo prenylation will help define the prenylated proteome and identify prenylation‐dependent pathways for therapeutic targeting.