Specific labeling of isoprenylated proteins: application to study inhibitors of the post-translational farnesylation and geranylgeranylation.
Specific labeling of either farnesylated or geranylgeranylated proteins in human PC-3 prostate cancer cell line was obtained by suppression of mevalonic acid biosynthesis with lovastatin, 50 microM, followed by supplementation of cell culture medium with either [3H]farnesyl- or [3H]geranylgeranyl-pyrophosphate. The immunoprecipitation of either a farnesylated (p21 ras) or geranylgeranylated (p21 rap 1) protein demonstrated that labeling was specific since proteins were detected only if the appropriate isoprenoid was added to the culture medium. TLC analysis indicated that no conversion of one isoprenoid to the other occurred in these conditions. The selective labeling of either farnesylated or geranylgeranylated proteins may be a valuable tool for the development of inhibitors of isoprenoid transferases as a potential new class of antitumor agents.
Danesi
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Division of Hematology/Oncology, University of Virginia, Charlottesville 22908.
McLellan
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CA
Myers
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CE
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008148:Lovastatin / Q000494:pharmacology; D008297:Male; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D011471:Prostatic Neoplasms; D017368:Protein Prenylation; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011865:Radioisotope Dilution Technique; D012717:Sesquiterpenes; D014316:Tritium; D014407:Tumor Cells, Cultured
D009363:Neoplasm Proteins; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D014316:Tritium; C004808:farnesyl pyrophosphate; D008148:Lovastatin; C002963:geranylgeranyl pyrophosphate