The importance of the post-translational lipid modifications farnesylation and geranylgeranylation in protein localization and function coupled with the critical role of prenylated proteins in malignant transformation has prompted interest in their biology and the development of farnesyl transferase and geranylgeranyl transferase inhibitors (FTIs and GGTIs) as chemical probes and anticancer agents. The ability to measure protein prenylation before and after FTI and GGTI treatment is important to understanding and interpreting the effects of these agents on signal transduction pathways and cellular phenotypes, as well as to the use of prenylation as a biomarker. Here we describe protocols to measure the degree of protein prenylation by farnesyl transferase or geranylgeranyl transferase in vitro, in cultured cells and in tumors from animals and humans. The assays use [3H]farnesyl diphosphate and [3H]geranylgeranyl diphosphate, electrophoretic mobility shift, membrane association using subcellular fractionation or immunofluorescence of intact cells, [3H]mevalonic acid labeling, followed by immunoprecipitation and SDS-PAGE, and in vitro transcription, translation and prenylation in reticulocyte lysates. These protocols require from 1 d (enzyme assays) to up to 3 months (autoradiography of [3H]-labeled proteins).
This is a preview of subscription content, access via your institution
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Hancock, J.F., Magee, A.I., Childs, J.E. & Marshall, C.J. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell 57, 1167–1177 (1989).
Casey, P.J., Solski, P.A., Der, C.J. & Buss, J.E. p21ras is modified by a farnesyl isoprenoid. Proc. Natl. Acad. Sci. USA 86, 8323–8327 (1989).
Falsetti, S.C. et al. Geranylgeranyltransferase I inhibitors target RalB to inhibit anchorage-dependent growth and induce apoptosis, and RalA to inhibit anchorage-independent growth. Mol. Cell. Biol. 27, 8003–8014 (2007).
Wang, C. et al. Identification of FBL2 as a geranylgeranylated cellular protein required for hepatitis C virus RNA replication. Mol. Cell 18, 425–434 (2005).
Kohl, N.E. et al. Protein farnesyltransferase inhibitors block the growth of ras-dependent tumors in nude mice. Proc. Natl. Acad. Sci. USA 91, 9141–9145 (1994).
Bishop, W.R. et al. Novel tricyclic inhibitors of farnesyl protein transferase. Biochemical characterization and inhibition of Ras modification in transfected Cos cells. J. Biol. Chem. 270, 30611–30618 (1995).
Sun, J. et al. Antitumor efficacy of a novel class of non-thiol-containing peptidomimetic inhibitors of farnesyltransferase and geranylgeranyltransferase I: combination therapy with the cytotoxic agents cisplatin, taxol, and gemcitabine. Cancer Res. 59, 4919–4926 (1999).
End, D.W. et al. Characterization of the antitumor effects of the selective farnesyl protein transferase inhibitor R115777 in vivo and in vitro. Cancer Res. 61, 131–137 (2001).
Sepp-Lorenzino, L. et al. A peptidomimetic inhibitor of farnesyl:protein transferase blocks the anchorage-dependent and -independent growth of human tumor cell lines. Cancer Res. 55, 5302–5309 (1995).
Zheng, H. et al. Ras homologue enriched in brain is a critical target of farnesyltransferase inhibitors in non-small cell lung cancer cells. Cancer Lett. 297, 117–125 (2010).
Sun, J., Qian, Y., Hamilton, A.D. & Sebti, S.M. Ras CAAX peptidomimetic FTI 276 selectively blocks tumor growth in nude mice of a human lung carcinoma with K-Ras mutation and p53 deletion. Cancer Res. 55, 4243–4247 (1995).
Mangues, R. et al. Antitumor effect of a farnesyl protein transferase inhibitor in mammary and lymphoid tumors overexpressing N-ras in transgenic mice. Cancer Res. 58, 1253–1259 (1998).
Cohen-Jonathan, E. et al. The farnesyltransferase inhibitor L744,832 reduces hypoxia in tumors expressing activated H-ras. Cancer Res. 61, 2289–2293 (2001).
Du, W., Lebowitz, P.F. & Prendergast, G.C. Cell growth inhibition by farnesyltransferase inhibitors is mediated by gain of geranylgeranylated RhoB. Mol. Cell. Biol. 19, 1831–1840 (1999).
Han, J.Y. et al. Hypoxia-inducible factor 1alpha and antiangiogenic activity of farnesyltransferase inhibitor SCH66336 in human aerodigestive tract cancer. J. Natl. Cancer Inst. 97, 1272–1286 (2005).
Kim, C.K. et al. The farnesyltransferase inhibitor LB42708 suppresses vascular endothelial growth factor-induced angiogenesis by inhibiting ras-dependent mitogen-activated protein kinase and phosphatidylinositol 3-kinase/Akt signal pathways. Mol. Pharmacol. 78, 142–150 (2010).
Sun, S.Y., Zhou, Z., Wang, R., Fu, H. & Khuri, F.R. The farnesyltransferase inhibitor Lonafarnib induces growth arrest or apoptosis of human lung cancer cells without downregulation of Akt. Cancer Biol. Ther. 3, 1092–1098 discussion 1099–1101 (2004).
Kazi, A. et al. Blockade of protein geranylgeranylation inhibits Cdk2-dependent p27Kip1 phosphorylation on Thr187 and accumulates p27Kip1 in the nucleus: implications for breast cancer therapy. Mol. Cell. Biol. 29, 2254–2263 (2009).
Moasser, M.M. et al. Farnesyl transferase inhibitors cause enhanced mitotic sensitivity to taxol and epothilones. Proc. Natl. Acad. Sci. USA 95, 1369–1374 (1998).
Hoover, R.R., Mahon, F.X., Melo, J.V. & Daley, G.Q. Overcoming STI571 resistance with the farnesyl transferase inhibitor SCH66336. Blood 100, 1068–1071 (2002).
Zhu, K. et al. Farnesyltransferase inhibitor R115777 (Zarnestra, tipifarnib) synergizes with paclitaxel to induce apoptosis and mitotic arrest to inhibit tumor growth of multiple myeloma cells. Blood 105, 4759–4766 (2005).
Punt, C.J., van Maanen, L., Bol, C.J., Seifert, W.F. & Wagener, D.J. Phase I and pharmacokinetic study of the orally administered farnesyl transferase inhibitor R115777 in patients with advanced solid tumors. Anticancer Drugs 12, 193–197 (2001).
Castro, A.F., Rebhun, J.F., Clark, G.J. & Qilliam, L.A. Rheb binds tuberous sclerosis complex 2 (TSC2) and promotes S6 kinase activation in a rapamycin- and farnesylation-dependent manner. J. Biol. Chem. 278, 32493–32496 (2003).
Mavrakis, K. et al. Tumorigenic activity and therapeutic inhibition of Rheb GTPase. Genes Dev. 22, 2178–2188 (2008).
Caruso, M.G. et al. Increased farnesyltransferase activity in human colorectal cancer: relationship with clinicopathological features and K-ras mutation. Scand. J. Gastroenterol. 38, 80–85 (2003).
Ryan, D.P. et al. Phase I clinical trial of the farnesyltransferase inhibitor BMS-214662 given as a 1-hour intravenous infusion in patients with advanced solid tumors. Clin. Cancer Res. 10, 2222–2230 (2004).
Kurzrock, R. et al. Phase I study of alternate-week administration of tipifarnib in patients with myelodysplastic syndrome. Clin. Cancer Res. 14, 509–514 (2008).
Kurzrock, R. et al. Farnesyltransferase inhibitor R115777 in myelodysplastic syndrome: clinical and biologic activities in the phase 1 setting. Blood 102, 4527–4534 (2003).
Adjei, A.A. et al. Phase II study of the farnesyl transferase inhibitor R115777 in patients with advanced non-small-cell lung cancer. J. Clin. Oncol. 21, 1760–1766 (2003).
Sparano, J.A. et al. Phase II trial of tipifarnib plus neoadjuvant doxorubicin-cyclophosphamide in patients with clinical stage IIB-IIIC breast cancer. Clin. Cancer Res. 15, 2942–2948 (2009).
Zhang, F.L. & Casey, P.J. Protein prenylation: molecular mechanisms and functional consequences. Annu. Rev. Biochem. 65, 241–269 (1996).
Lane, K.T. & Beese, L.S. Structural biology of protein farnesyltransferase and geranylgeranyltransferase type I. J. Lipid Res. 47, 681–699 (2006).
Perez-Sala, D. Protein isoprenylation in biology and disease: General overview and perspectives from studies with genetically engineered animals. Front. Biosci. 12, 4456–4472 (2007).
Carboni, J.M. et al. Farnesyltransferase inhibitors are inhibitors of Ras but not R-Ras2/TC21, transformation. Oncogene 10, 1905–1913 (1995).
Moores, S.L. et al. Sequence dependence of protein isoprenylation. J. Biol. Chem. 266, 14603–14610 (1991).
Roskoski, R. Jr. & Ritchie, P. Role of the carboxyterminal residue in peptide binding to protein farnesyltransferase and protein geranylgeranyltransferase. Arch. Biochem. Biophys. 356, 167–176 (1998).
Boutin, J.A. et al. Chromatographic assay and peptide substrate characterization of partially purified farnesyl- and geranylgeranyltransferases from rat brain cytosol. Arch. Biochem. Biophys. 354, 83–94 (1998).
Reid, T.S., Terry, K.L., Casey, P.J. & Beese, L.S. Crystallographic analysis of CaaX prenyltransferases complexed with substrates defines rules of protein substate selectivity. J. Mol. Biol. 343, 417–433 (2004).
Whyte, D.B. et al. K- and N-Ras are geranylgeranylated in cells treated with farnesyl protein transferase inhibitors. J. Biol. Chem. 272, 14459–14464 (1997).
Winter-Vann, A.M. & Casey, P.J. Post-prenylation-processing enzymes as new targets in oncogenesis. Nat. Rev. Cancer 5, 405–412 (2005).
Linder, M.E. & Deschenes, R.J. Palmitoylation: policing protein stability and traffic. Nat. Rev. Mol. Cell Biol. 8, 74–84 (2007).
Baekkeskov, S. & Kanaani, J. Palmitoylation cycles and regulation of protein function (Review). Mol. Membr. Biol. 26, 42–54 (2009).
Maurer-Stroh, S. & Eisenhaber, F. Refinement and prediction of protein prenylation motifs. Genome Biol. 6, R55 (2005).
Maurer-Stroh, S. et al. Towards complete sets of farnesylated and geranylgeranylated proteins. PLoS Comput. Biol. 3, e66 (2007).
Sebti, S.M. Protein farnesylation: implications for normal physiology, malignant transformation, and cancer therapy. Cancer Cell 7, 297–300 (2005).
Mijimolle, N. et al. Protein farnesyltransferase in embryogenesis, adult homeostasis, and tumor development. Cancer Cell 7, 313–324 (2005).
Lerner, E.C. et al. Ras CAAX peptidomimetic FTI-277 selectively blocks oncogenic ras signaling by inducing cytoplasmic accumulation of inactive ras-raf complexes. J. Biol. Chem. 270, 26802–26806 (1995).
Willumsen, B.M., Christensen, A., Hubbert, N.L., Papageorge, A.G. & Lowy, D.R. The p21 ras C-terminus is required for transformation and membrane association. Nature 310, 583–586 (1984).
Yang, S.H. et al. Caution! Analyze transcripts from conditional knockout alleles. Transgenic Res. 18, 483–489 (2009).
Liu, M. et al. Targeting the protein prenyltransferases efficiently reduces tumor development in mice with K-RAS-induced lung cancer. Proc. Natl. Acad. Sci. USA 107, 6471–6476 (2010).
Willumsen, B.M., Norris, K., Papageorge, A.G., Hubbert, N.L. & Lowy, D.R. Harvey murine sarcoma virus p21 ras protein: biological and biochemical significance of the cysteine nearest the carboxy terminus. EMBO J. 3, 2581–2585 (1984).
Butrynski, J.E., Jones, T.L., Backlund, P.S. Jr. & Spiegel, A.M. Differential isoprenylation of carboxy-terminal mutants of an inhibitory G-protein alpha-subunit: neither farnesylation nor geranylgeranylation is sufficient for membrane attachment. Biochemistry 31, 8030–8035 (1992).
Ohya, Y. et al. Yeast CAL1 is a structural and functional homologue to the DPR1 (RAM) gene involved in ras processing. J. Biol. Chem. 266, 12356–12360 (1991).
Therrien, M. et al. KSR, a novel protein kinase required for RAS signal transduction. Cell 83, 879–888 (1995).
Jackson, J.H. et al. Farnesol modification of Kirsten-ras exon 4B protein is essential for transformation. Proc. Natl. Acad. Sci. USA 87, 3042–3046 (1990).
Campbell, P.M. & Der, C.J. Oncogenic Ras and its role in tumor cell invasion and metastasis. Semin. Cancer Biol. 14, 105–114 (2004).
Stokoe, D., Macdonald, S.G., Cadwallader, K., Symons, M. & Hancock, J.F. Activation of Raf as a result of recruitment to the plasma membrane. Science 264, 1463–1467 (1994).
Rodriguez-Viciana, P. et al. Phosphatidylinositol-3-OH kinase as a direct target of Ras. Nature 370, 527–532 (1994).
Appels, N.M.G.M., Beijnen, J.H. & Schellens, J.H.M. Development of farnesyltransferase inhibitors: a review. Oncologist 10, 565–578 (2005).
Karnoub, A.E. & Weinberg, R.A. Ras oncogenes: split personalities. Nat. Rev. Mol. Cell Biol. 9, 517–531 (2008).
Sun, J., Qian, Y., Hamilton, A.D. & Sebti, S.M. Both farnesyltransferase and geranylgeranyltransferase I inhibitors are required for inhibition of oncogenic K-Ras prenylation but each alone is sufficient to suppress human tumor growth in nude mouse xenografts. Oncogene 16, 1467–1473 (1998).
Kohl, N.E. et al. Inhibition of farnesyltransferase induces regression of mammary salivary carcinomas in ras transgenic mice. Nat. Med. 1, 792–797 (1995).
Omer, C.A. et al. Mouse mammary tumor virus-Ki-rasB transgenic mice develop mammary carcinomas that can be growth-inhibited by a farnesyl:protein transferase inhibitor. Cancer Res. 60, 2680–2688 (2000).
Sebti, S.M. & Der, C.J. Searching for the elusive targets of farnesyltransferase inhibitors. Nat. Rev. Cancer 3, 945–951 (2003).
Basso, A.D., Kirschmeier, P. & Bishop, W.R. Lipid posttranslational modifications: farnesyl transferase inhibitors. J. Lipid Res. 47, 15–31 (2006).
Ashar, H.R. et al. Farnesyl transferase inhibitors block the farnesylation of CENP-E and CENP-F and alter the association of CENP-E with the microtubules. J. Biol. Chem. 275, 30451–30457 (2000).
Crespo, N.C., Ohkanda, J., Yen, T.J., Hamilton, A.D. & Sebti, S.M. The farnesyltransferase inhibitor, FTI-2153, blocks bipolar spindle formation and chromosome alignment and causes prometaphase accumulation during mitosis of human lung cancer cells. J. Biol. Chem. 276, 16161–16167 (2001).
Crespo, N.C. et al. The farnesyltransferase inhibitor, FTI-2153, inhibits bipolar spindle formation during mitosis independently of transformation and Ras and p53 mutation status. Cell Death Differ. 9, 702–709 (2002).
Sepp-Lorenzino, L. & Rosen, N. A farnesyl-protein transferase inhibitor induces p21 expression and G1 block in p53 wild type tumor cells. J. Biol. Chem. 273, 20243–20251 (1998).
Prendergast, G.C. Actin' up: RhoB in cancer and apoptosis. Nat. Rev. Cancer 1, 162–168 (2001).
Chen, Z. et al. Both farnesylated and geranylgeranylated RhoB inhibit malignant transformation and suppress human tumor growth in nude mice. J. Biol. Chem. 275, 17974–17978 (2000).
Van Cutsem, E. et al. Phase III trial of gemcitabine plus tipifarnib compared with gemcitabine plus placebo in advanced pancreatic cancer. J. Clin. Oncol. 22, 1430–1438 (2004).
Rao, S. et al. Phase III double-blind placebo-controlled study of farnesyl transferase inhibitor R115777 in patients with refractory advanced colorectal cancer. J. Clin. Oncol. 22, 3950–3957 (2004).
Harousseau, J.L. et al. A randomized phase 3 study of tipifarnib compared with best supportive care, including hydroxyurea, in the treatment of newly diagnosed acute myeloid leukemia in patients 70 years or older. Blood 114, 1166–1173 (2009).
Sparano, J.A. et al. Targeted inhibition of farnesyltransferase in locally advanced breast cancer: A phase I and II trial of Tipifarnib plus dose-dense doxirubicin and cyclophosphamide. J. Clin. Oncol. 24, 3013–3018 (2006).
Hamad, N.M. et al. Distinct requirements for Ras oncogenesis in human versus mouse cells. Genes Dev. 16, 2045–2057 (2002).
Lim, K.-H. et al. Activation of RalA is critical for Ras-induced tumorigenesis of human cells. Cancer Cell 7, 533–545 (2005).
Vogt, A., Sun, J.Z., Qian, Y.M., Hamilton, A.D. & Sebti, S.M. The geranylgeranyltransferase-I inhibitor GGTI-298 arrests human tumor cells in G0/G1 and induces p21WAF1/CIP1/SDI1 in a p53-independent manner. J. Biol. Chem. 272, 27224–27229 (1997).
Sun, J. et al. The geranylgeranyltransferase I inhibitor GGTI-298 induces hypophosphorylation of retinoblastoma and partner switching of cyclin-dependent kinase inhibitors. A potential mechanism for GGTI-298 antitumor activity. J. Biol. Chem. 274, 6930–6934 (1999).
Sun, J. et al. Geranylgeranyltransferase I inhibitor GGTI-2154 induces breast carcinoma apoptosis and tumor regression in H-ras transgenic mice. Cancer Res. 63, 8922–8929 (2003).
Dan, H.C. et al. Phosphatidylinositol-3-OH kinase/AKT and survivin pathways as critical targets for geranylgeranyltransferase I inhibitor-induced apoptosis. Oncogene 22, 706–715 (2004).
Sjogren, A.-K. et al. GGTase-1 deficiency reduces tumor formation and improves survival in mice with K-Ras-induced lung cancer. J. Clin. Invest. 117, 1294–1304 (2007).
O'Dwyer, P.J., Gallagher, M., Nguyen, B., Waddell, M.J. & Chiorean, E.G. Phase I accelerated dose-escalating safety and pharmacokinetic (PK) study of GGTI-2418, a novel geranylgeranyltransferase I inhibitor in patients with refractory solid tumors. Ann. Oncol. 21 (suppl 2), ii42 (2010).
Vogt, A. et al. Burkitt lymphoma Daudi cells contain two distinct farnesyltransferases with different divalent cation requirements. Biochemistry 34, 12398–12403 (1995).
Nigam, M., Seong, C.M., Qian, Y., Hamilton, A.D. & Sebti, S.M. Potent inhibition of human tumor p21ras farnesyltransferase by A1A2-lacking p21ras CA1A2X peptidomimetics. J. Biol. Chem. 268, 20695–20698 (1993).
Qian, Y. et al. Design and structural requirements of potent peptidomimetic inhibitors of p21ras farnesyltransferase. J. Biol. Chem. 269, 12410–12413 (1994).
Vogt, A. et al. A non-peptide mimetic of Ras-CAAX: selective inhibition of farnesyltransferase and Ras processing. J. Biol. Chem. 270, 660–664 (1995).
McGuire, T.F. et al. CAAX peptidomimetic FTI-244 decreases platelet-derived growth factor receptor tyrosine phosphorylation levels and inhibits stimulation of phosphatidylinositol 3-kinase but not mitogen-activated protein kinase. Biochem. Biophys. Res. Commun. 214, 295–303 (1995).
Lerner, E.C., Qian, Y., Hamilton, A.D. & Sebti, S.M. Disruption of oncogenic K-Ras4B processing and signaling by a potent geranylgeranyltransferase I inhibitor. J. Biol. Chem. 270, 26770–26773 (1995).
Hunt, J.T. et al. Discovery of (R)-7-cyano-2,3,4, 5-tetrahydro-1-(1H-imidazol-4-ylmethyl)-3- (phenylmethyl)-4-(2-thienylsulfonyl)-1H-1,4-benzodiazepine (BMS-214662), a farnesyltransferase inhibitor with potent preclinical antitumor activity. J. Med. Chem. 43, 3587–3595 (2000).
Cox, A.D., Hisaka, M.M., Buss, J.E. & Der, C.J. Specific isoprenoid modification is required for function of normal, but not oncogenic, Ras protein. Mol. Cell. Biol. 12, 2606–2615 (1992).
Sinensky, M., Fantle, K. & Dalton, M. An antibody which specifically recognizes prelamin A but not mature lamin A: application to detection of blocks in farnesylation-dependent protein processing. Cancer Res. 54, 3229–3232 (1994).
Lerner, E.C. et al. Inhibition of the prenylation of K-Ras, but not H- or N-Ras, is highly resistant to CAAX peptidomimetics and requires both a farnesyltransferase and geranylgeranyltransferase I inhibitor in human tumor cell lines. Oncogene 15, 1283–1288 (1997).
Vogt, A., Qian, Y., McGuire, T.F., Hamilton, A.D. & Sebti, S.M. Protein geranylgeranylation, not farnesylation, is required for the G1 to S phase transition in mouse fibroblasts. Oncogene 13, 1991–1999 (1996).
Delarue, F.L. et al. Farnesyltransferase and geranylgeranyltransferase I inhibitors upregulate RhoB expression by HDAC1 dissociation, HAT association and histone acetylation of the RhoB promoter. Oncogene 26, 633–640 (2007).
Brunner, T.B. et al. Pancreatic cancer cell radiation survival and prenyltransferase inhibition: the role of K-Ras. Cancer Res. 65, 8433–8441 (2005).
Wang, D.-A. & Sebti, S.M. Palmitoylated cysteine 192 is required for RhoB tumor-suppressive and apoptotic activities. J. Biol. Chem. 280, 19243–19249 (2005).
Bivona, T.G. et al. PKC regulates a farnesyl-electrostatic switch on K-ras that promotes its association with Bcl-Xl on mitochondria and induces apoptosis. Mol. Cell 21, 481–493 (2006).
Bivona, T.G., Quatela, S. & Philips, M.R. Analysis of Ras activation in living cells with GFP-RBD. Methods Enzymol. 407, 128–143 (2006).
Benetka, W., Koranda, M., Maurer-Stroh, S., Pittner, F. & Eisenhaber, F. Farnesylation or geranylgeranylation? Efficient assays for testing protein prenylation in vitro and in vivo. BMC Biochem. 7, 6 (2006).
Kho, Y. et al. A tagging-via-substrate technology for detection and proteomics of farnesylated proteins. Proc. Natl. Acad. Sci. USA 101, 12479–12484 (2004).
Onono, F.O. et al. A tagging-via-substrate approach to detect the farnesylated proteome using two-dimensional electrophoresis coupled with Western blotting. Mol. Cell. Proteomics 9, 742–751 (2010).
Nguyen, U.T. et al. Analysis of the eukaryotic prenylome by isoprenoid affinity tagging. Nat. Chem. Biol. 5, 227–235 (2009).
Troutman, J.M., Roberts, M.J., Andres, D.A. & Spielmann, H.P. Tools to analyze protein farnesylation in cells. Bioconjug. Chem. 16, 1209–1217 (2005).
Chan, L.N. et al. A novel approach to tag and identify geranylgeranylated proteins. Electrophoresis 30, 3598–3606 (2009).
Degraw, A.J. et al. Evaluation of alkyne-modified isoprenoids as chemical reporters of protein prenylation. Chem. Biol. Drug Des. 6, 460–471 (2010).
Hannoush, R.N. & Sun, J. The chemical toolbox for monitoring protein fatty acylation and prenylation. Nat. Chem. Biol. 6, 498–506 (2010).
Kita, T., Brown, M.S. & Goldstein, J.L. Feedback regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in livers of mice treated with mevinolin, a competitive inhibitor of the reductase. J. Clin. Invest. 66, 1094–1100 (1980).
Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976).
Berndt, N. et al. The Akt activation inhibitor TCN-P inhibits Akt phosphorylation by binding to the PH domain of Akt and blocking its recruitment to the plasma membrane. Cell Death Differ. 17, 1795–1804 (2010).
This work was supported by grants from the NIH CA067771 (S.M.S) and CA098473 (S.M.S.). We thank A.D. Hamilton and his group for a wonderful and highly productive collaboration and M.A. Blaskovich and A. Kazi for their excellent suggestions. We also thank the past and present members of the Sebti lab for their contributions to this work, particularly J. Adnane, M.E. Balasis, M.A. Blaskovich, C. Bucher, P.M. Campbell, A.E. Carie, Z. Chen, N.C.Crespo, F. Delarue, S.C. Falsetti, K. Forinash, K. Jiang, A. Kazi, E.C. Lerner, T.F. McGuire, M. Nigam, S. Paquette, R. Patel, J. Sun, Y. Sun, V. Thai, A. Vogt, D. Wang, A. Tecleab, H. Yang and K. Zhu.
The authors declare no competing financial interests.
About this article
Cite this article
Berndt, N., Sebti, S. Measurement of protein farnesylation and geranylgeranylation in vitro, in cultured cells and in biopsies, and the effects of prenyl transferase inhibitors. Nat Protoc 6, 1775–1791 (2011). https://doi.org/10.1038/nprot.2011.387
This article is cited by
Ubiquitin-like protein 3 (UBL3) is required for MARCH ubiquitination of major histocompatibility complex class II and CD86
Nature Communications (2022)
Protein farnesylation is upregulated in Alzheimer’s human brains and neuron-specific suppression of farnesyltransferase mitigates pathogenic processes in Alzheimer’s model mice
Acta Neuropathologica Communications (2021)
Metabolic labeling with an alkyne probe reveals similarities and differences in the prenylomes of several brain-derived cell lines and primary cells
Scientific Reports (2021)
BMC Cancer (2020)
Scientific Reports (2020)