Evaluation of alkyne-modified isoprenoids as chemical reporters of protein prenylation

摘要: Protein prenyltransferases catalyze the attachment of C15 (farnesyl) and C20 (geranylgeranyl) groups to proteins at specific sequences localized or near C-termini proteins. Determination protein prenyltransferase substrates affected by inhibition these enzymes is critical for enhancing knowledge mechanism such potential drugs. Here, we investigate utility alkyne-containing isoprenoid analogs chemical proteomics experiments showing that compounds readily penetrate mammalian cells in culture become incorporated into are normally prenylated. Derivatization via Cu(I) catalyzed click reaction with a fluorescent azide reagent allows be visualized their relative levels analyzed. Simultaneous treatment probes inhibitors prenylation reveals decreases some but not all labeled Two-dimensional electrophoretic separation followed mass spectrometric analysis allowed several unambiguously identified. Docking density functional theory calculations suggest substrate specificity farnesyl transferase may vary depending on whether azide- alkyne-based employed. These results demonstrate proteomic applications.