Abstract
In this article, assays on the analytical determination of farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), two important isoprenoid intermediates at biochemically relevant branching points in the mevalonate pathway, are summarized and reviewed. There is considerable recent interest in the measurement of these two isoprenoids because of their direct involvement in several diseases, for example, statins lower cholesterol by inhibiting 3-hydroxy-3-methylglutaryl-CoA reductase but equally affect other metabolite biosyntheses. The isoprenoids FPP and GGPP are key intermediates due to their role as CaaX-specific substrates for posttranslational modification of proteins (protein prenylation). Disease pathologies and therapeutic efficacy of different treatments (e.g., cholesterol-lowering drugs) may lead to a reduction in isoprenoid levels and an accompanying reduction in prenylation of specific proteins. To understand the exact biochemical role of the isoprenoids FPP and GGPP, we need to know their levels. Several recent studies have shown exact levels of FPP and GGP in plasma and relevant tissues and their modulation following treatment. Furthermore, by directly measuring the extent of protein prenylation and identifying target proteins, further insight into the exact biochemical nature of the pathology and regulatory mechanisms will be possible. This short review aims to highlight the relevant literature on the analytical determination of the free isoprenoids FPP and GGPP in biological tissue as well as techniques for directly measuring prenylated proteins.
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References
Holstein SA, Hohl RJ (2004) Isoprenoids: remarkable diversity of form and function. Lipids 39:293
McTaggart SJ (2006) Isoprenylated proteins. Cell Mol Life Sci 63:255
Rohmer M (2003) Mevalonate-independent methylerythritol phosphate pathway for isoprenoid biosynthesis. Elucidation and distribution. Pure Appl Chem 75:375
Eisenreich W, Bacher A, Arigoni D, Rohdich F (2004) Biosynthesis of isoprenoids via the non-mevalonate pathway. Cell Mol Life Sci 61:1401
Goldstein JL, Brown MS (1990) Regulation of the mevalonate pathway. Nature 343:425
Buhaescu I, Izzedine H (2007) Mevalonate pathway: a review of clinical and therapeutical implications. Clin Biochem 40:575
Harris CM, Poulter CD (2000) Recent studies of the mechanism of protein prenylation. Nat Prod Rep 17:137
Winter-Vann AM, Casey PJ (2005) Post-prenylation-processing enzymes as new targets in oncogenesis. Nat Rev Cancer 5:405
Zhang FL, Casey PJ (1996) Protein prenylation: molecular mechanisms and functional consequences. Annu Rev Biochem 65:241
Gelb MH, Scholten JD, Sebolt-Leopold JS (1998) Protein prenylation: from discovery to prospects for cancer treatment. Curr Opin Chem Biol 2:40
Auer J, Berent R, Weber T, Eber B (2002) Clinical significance of pleiotropic effects of statins: lipid reduction and beyond. Curr Med Chem 9:1831
Fonseca ACRG, Resende R, Oliveira CR, Pereira CMF (2010) Cholesterol and statins in Alzheimer's disease: current controversies. Exp Neurol 223:282
Swanson KM, Hohl RJ (2006) Anti-cancer therapy: targeting the mevalonate pathway. Curr Cancer Drug Tar 6:15
Wolozin B, Kellman W, Ruosseau P, Celesia GG, Siegel G (2000) Decreased prevalence of Alzheimer disease associated with 3-hydroxy-3-methyglutaryl coenzyme A reductase inhibitors. Arch Neurol 57:1439
Jick H, Zornberg GL, Jick SS, Seshadri S, Drachman DA (2000) Statins and the risk of dementia. Lancet 356:1627
Ostrowski SM, Wilkinson BL, Golde TE, Landreth G (2007) Statins reduce amyloid-beta production through inhibition of protein isoprenylation. J Biol Chem 282:26832
Hooff GP, Wood WG, Müller WE, Eckert GP (2010) Isoprenoids, small GTPases and Alzheimer's disease. Biochim Biophys Acta 1801:896
Cole SL, Vassar R (2006) Isoprenoids and Alzheimer's disease: a complex relationship. Neurobiol Dis 22:209
Mijimolle N, Velasco J, Dubus P, Guerra C, Weinbaum CA, Casey PJ, Campuzano V, Barbacid M (2005) Protein farnesyltransferase in embryogenesis, adult homeostasis, and tumor development. Cancer Cell 7:313
Sebti SM (2005) Protein farnesylation: implications for normal physiology, malignant transformation, and cancer therapy. Cancer Cell 7:297
Ferri N, Paoletti R, Corsini A (2005) Lipid-modified proteins as biomarkers for cardiovascular disease: a review. Biomarkers 10:219
Greenwood J, Steinman L, Zamvil SS (2006) Statin therapy and autoimmune disease: from protein prenylation to immunomodulation. Nat Rev Immunol 6:358
Houten SM, Frenkel J, Waterham HR (2003) Isoprenoid biosynthesis in hereditary periodic fever syndromes and inflammation. Cell Mol Life Sci 60:1118
Weivoda MM, Hohl RJ (2011) Effects of farnesyl pyrophosphate accumulation on calvarial osteoblast differentiation. Endocrinol (in press)
Gelb MH, Brunsveld L, Hrycyna CA, Michaelis S, Tamanoi F, Van Voorhis WC, Waldmann H (2006) Therapeutic intervention based on protein prenylation and associated modifications. Nat Chem Biol 2:518
Liao JK (2002) Isoprenoids as mediators of the biological effects of statins. J Clin Invest 110:285
Dugan RE, Rasson E, Porter JW (1968) Separation of water-soluble steroid and carotenoid precursors by DEAE-cellulose column chromatography. Anal Biochem 22:249
Bruenger E, Rilling HC (1988) Determination of isopentenyl diphosphate and farnesyl diphosphate in tissue samples with a comment on secondary regulation of polyisoprenoid biosynthesis. Anal Biochem 173:321
Zhang D, Poulter CD (1993) Analysis and purification of phosphorylated isoprenoids by reversed-phase HPLC. Anal Biochem 213:356
Keller R, Bangalore P, Robert J, Swanson M (1996) Analysis of isoprenoid phosphates and oligophosphates by capillary zone electrophoresis. J Chromatogr A 737:325
Andersson P, Pfeffer W, Blomberg L (1995) Indirect detection in capillary electrophoresis comparison between indirect UV and indirect laser-induced fluorescence detection for the determination of isoprenyl pyrophosphates. J Chromatogr A 699:323
Davisson VJ, Sharp TR, Poulter CD (1988) Negative ion fast atom bombardment mass spectrometry of isoprenoid diphosphates and related analogs. Bioorg Chem 16:111
Kitaoka M, Nagaki H, Kinoshita T, Kurabayashi M, Koyama T, Ogura K (1990) Negative ion fast atom bombardment-tandem mass spectrometry for structural analysis of isoprenoid diphosphates. Anal Biochem 185:182
Henneman L, van Cruchten AG, Denis SW, Amolins MW, Placzek AT, Gibbs RA, Kulik W, Waterham HR (2008) Detection of nonsterol isoprenoids by HPLC-MS/MS. Anal Biochem 383:18
Henneman L, van Cruchten AG, Kulik W, Waterham HR (2011) Inhibition of the isoprenoid biosynthesis pathway; detection of intermediates by UPLC-MS/MS. Biochim Biophys Acta 1811:227
Lee SH, Raboune S, Walker JM, Bradshaw HB (2010) Distribution of endogenous farnesyl pyrophosphate and four species of lysophosphatidic acid in rodent brain. Int J Mol Sci 11:3965
Song L (2003) Detection of farnesyl diphosphate accumulation in yeast ERG9 mutants. Anal Biochem 317:180
Vallon T, Ghanegaonkar S, Vielhauer O, Müller A, Albermann C, Sprenger G, Reuss M, Lemuth K (2008) Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains. Appl Microbiol Biotechnol 81:175
D'Alexandri FL, Gozzo FC, Eberlin MN, Katzin AM (2006) Electrospray ionization mass spectrometry analysis of polyisoprenoid alcohols via Li+ cationization. Anal Biochem 355:189
Teshima K, Kondo T (2008) Analytical method for determination of allylic isoprenols in rat tissues by liquid chromatography/tandem mass spectrometry following chemical derivatization with 3-nitrophtalic anhydride. J Pharm Biomed Anal 47:560
McCaskill D, Croteau R (1993) Procedures for the isolation and quantification of the intermediates of the mevalonic acid pathway. Anal Biochem 215:142
Lutz RJ, McLain TM, Sinensky M (1992) Feedback inhibition of polyisoprenyl pyrophosphate synthesis from mevalonate in vitro. Implications for protein prenylation. J Biol Chem 267:7983
Yang T, Zabriskie TM, Poulter CD (1984) Investigation of isoprenoid benzoates and naphthoates by reversed-phase liquid chromatography isocratic elution characteristics of benzoates and naphthoates of C5–C20 terpenoid alcohols. J Chromatogr A 312:121
Yang T, Zabriskie TM, Poulter CD (1985) Reversed-phase high-performance liquid chromatography of C5 to C20 isoprenoid benzoates and naphthoates. Methods Enzymol 111:252
Epstein WW, Wang Z, Leining LM, Lever DC (1996) Identification of prenylcysteines and prenylated proteins by formation of substituted naphthopyrans. J Org Chem 61:4890
Saisho Y, Morimoto A, Umeda T (1997) Determination of farnesyl pyrophosphate in dog and human plasma by high-performance liquid chromatography with fluorescence detection. Anal Biochem 252:89
Pompliano DL, Gomez RP, Anthony NJ (1992) Intramolecular fluorescence enhancement: a continuous assay of Ras farnesyl:protein transferase. J Am Chem Soc 114:7945
Pickett WC, Zhang FL, Silverstrim C, Schow SR, Wick MM, Kerwar SS (1995) A fluorescence assay for geranylgeranyl transferase type I. Anal Biochem 225:60
Tong H, Holstein SA, Hohl RJ (2005) Simultaneous determination of farnesyl and geranylgeranyl pyrophosphate levels in cultured cells. Anal Biochem 336:51
Tong H, Wiemer AJ, Neighbors JD, Hohl RJ (2008) Quantitative determination of farnesyl and geranylgeranyl diphosphate levels in mammalian tissue. Anal Biochem 378:138
Holstein SA, Tong H, Kuder CH, Hohl RJ (2009) Quantitative determination of geranyl diphosphate levels in cultured human cells. Lipids 44:1055
Holstein SA, Tong H, Hohl RJ (2010) Differential activities of thalidomide and isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells. Leuk Res 34:344
Hooff GP, Volmer DA, Wood WG, Müller WE, Eckert GP (2008) Isoprenoid quantitation in human brain tissue: a validated HPLC-fluorescence detection method for endogenous farnesyl- (FPP) and geranylgeranylpyrophosphate (GGPP). Anal Bioanal Chem 392:673
Murthy S, Tong H, Hohl RJ (2005) Regulation of fatty acid synthesis by farnesyl pyrophosphate. J Biol Chem 280:41793
Hooff GP, Patel N, Wood GW, Müller WE, Eckert GP, Volmer DA (2010) A rapid and sensitive assay for determining human brain levels of farnesyl-(FPP) and geranylgeranylpyrophosphate (GGPP) and transferase activities using UHPLC-MS/MS. Anal Bioanal Chem 398:1801
Hooff GP, Peters I, Wood WG, Müller WE, Eckert GP (2010) Modulation of cholesterol, farnesylpyrophosphate, and geranylgeranylpyrophosphate in neuroblastoma SH-SY5Y-APP695 cells: impact on amyloid beta-protein production. Mol Neurobiol 41:341
Eckert GP, Hooff GP, Strandjord DM, Igbavboa U, Volmer DA, Müller WE, Wood WG (2009) Regulation of the brain isoprenoids farnesyl- and geranylgeranylpyrophosphate is altered in male Alzheimer patients. Neurobiol Dis 35:251
Dudakovic A, Wiemer AJ, Lamb KM, Vonnahme LA, Dietz SE, Hohl RJ (2008) Inhibition of geranylgeranyl diphosphate synthase induces apoptosis through multiple mechanisms and displays synergy with inhibition of other isoprenoid biosynthetic enzymes. J Pharmacol Exp Ther 324:1028
Berezovski M, Li W, Poulter CD, Krylov SN (2002) Measuring the activity of farnesyltransferase by capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis 23:3398
Ishiguro H, Kawata S, Yamasaki E, Matsuda Y, Fujii S, Matsuzawa Y (1995) High-performance liquid chromatographic assay for farnesyl-protein transferase activity with dabsylated peptide. J Chromatogr B 663:35
Simonen M, Ibig-Rehm Y, Hofmann G, Zimmermann J, Albrecht G, Magnier M, Heidinger V, Gabriel D (2008) High-content assay to study protein prenylation. J Biomol Screen 13:456
Wollack JW, Zeliadt NA, Mullen DG, Amundson G, Geier S, Falkum S, Wattenberg EV, Barany G, Distefano MD (2009) Multifunctional prenylated peptides for live cell analysis. J Am Chem Soc 131:7293
Pais JE, Bowers KE, Stoddard AK, Fierke CA (2005) A continuous fluorescent assay for protein prenyltransferases measuring diphosphate release. Anal Biochem 345:302
Maltese WA, Erdman RA (1989) Characterization of isoprenoid involved in the post-translational modification of mammalian cell proteins. J Biol Chem 264:18168
Joly A, Popják G, Edwards PA (1991) In vitro identification of a soluble protein:geranylgeranyl transferase from rat tissues. J Biol Chem 266:13495
Epstein WW, Lever DC, Rilling HC (1990) Prenylated proteins: synthesis of geranylgeranylcysteine and identification of this thioether amino acid as a component of proteins in CHO cells. Proc Natl Acad Sci U S A 87:7352
Epstein WW, Lever D, Leining LM, Bruenger E, Rilling HC (1991) Quantitation of prenylcysteines by a selective cleavage reaction. Proc Natl Acad Sci U S A 88:9668
Hjertman M, Wejde J, Larsson O (2001) Characterization of hydrophobic prenyl groups of isoprenylated proteins in human cancer cells. Biochem Biophys Res Commun 288:736
Appels NMGM, Rosing H, Stephens TC, Schellens JHM, Beijnen JH (2006) Quantification of farnesylmethylcysteine in lysates of peripheral blood mononuclear cells using liquid chromatography coupled with electrospray tandem mass spectrometry: pharmacodynamic assay for farnesyl transferase inhibitors. Anal Chem 78:2617
Rilling HC, Bruenger E, Epstein WW, Crain PF (1990) Prenylated proteins: the structure of the isoprenoid group. Science 247:318
Farnsworth CC, Gelb MH, Glomset JA (1990) Identification of geranylgeranyl-modified proteins in HeLa cells. Science 247:320
Khosravi-Far R, Lutz RJ, Cox AD, Conroy L, Bourne JR, Sinensky M, Balch WE, Buss JE, Der CJ (1991) Isoprenoid modification of Rab proteins terminating in CC or CXC motifs. Proc Natl Acad Sci USA 88:6264
Kawata M, Farnsworth CC, Yoshida Y, Gelb MH, Glomset JA, Takai Y (1990) Posttranslationally processed structure of the human platelet protein smg p21B: evidence for geranylgeranylation and carboxyl methylation of the C-terminal cysteine. Proc Natl Acad Sci U S A 87:8960
Danesi R, McLellan CA, Myers CE (1995) Specific labeling of isoprenylated proteins: application to study inhibitors of the post-translational farnesylation and geranylgeranylation. Biochem Biophys Res Commun 206:637
Khan SG, Mukhtar H, Agarwal R (1995) A rapid and convenient filter-binding assay for ras p21 processing enzyme farnesyltransferase. J Biochem Biophys Methods 30:133
Benetka W, Koranda M, Maurer-Stroh S, Pittner F, Eisenhaber F (2006) Farnesylation or geranylgeranylation? Efficient assays for testing protein prenylation in vitro and in vivo. BMC Biochem 7:6
Nguyen UTT, Wu Y, Goodall A, Alexandrov K (2010) Analysis of protein prenylation in vitro and in vivo using functionalized phosphoisoprenoids. Curr Protoc Protein Sci 14:14.3
Dursina B, Reents R, Niculae A, Veligodsky A, Breitling R, Pyatkov K, Waldmann H, Goody RS, Alexandrov K (2005) A genetically encodable microtag for chemo-enzymatic derivatization and purification of recombinant proteins. Protein Expr Purif 39:71
Dursina B, Reents R, Delon C, Wu Y, Kulharia M, Thutewohl M, Veligodsky A, Kalinin A, Evstifeev V, Ciobanu D, Szedlacsek SE, Waldmann H, Goody RS, Alexandrov K (2006) Identification and specificity profiling of protein prenyltransferase inhibitors using new fluorescent phosphoisoprenoids. J Am Chem Soc 128:2822
Wu Y, Waldmann H, Reents R, Ebetino FH, Goody RS, Alexandrov K (2006) A protein fluorescence amplifier: continuous fluorometric assay for rab geranylgeranyltransferase. ChemBioChem 7:1859
Wu Y, Alexandrov K, Brunsveld L (2007) Synthesis of a fluorescent analogue of geranylgeranyl pyrophosphate and its use in a high-throughput fluorometric assay for Rab geranylgeranyltransferase. Nat Protoc 2:2704
Turek TC, Gaon I, Distefano MD, Strickland CL (2001) Synthesis of farnesyl diphosphate analogues containing ether-linked photoactive benzophenones and their application in studies of protein prenyltransferases. J Org Chem 66:3253
Liu X, Prestwich GD (2002) Didehydrogeranylgeranyl (ΔΔGG): a fluorescent probe for protein prenylation. J Am Chem Soc 124:20
Kho Y, Kim SC, Jiang C, Barma D, Kwon SW, Cheng J, Jaunbergs J, Weinbaum C, Tamanoi F, Falck J, Zhao Y (2004) A tagging-via-substrate technology for detection and proteomics of farnesylated proteins. Proc Natl Acad Sci U S A 101:12479
Köhn M, Breinbauer R (2004) The Staudinger ligation-a gift to chemical biology. Angew Chem Int Ed 43:3106
Bräse S, Gil C, Knepper K, Zimmermann V (2005) Organic azides: an exploding diversity of a unique class of compounds. Angew Chem Int Ed 44:5188
Rose MW, Rose ND, Boggs J, Lenevich S, Xu J, Barany G, Distefano MD (2005) Evaluation of geranylazide and farnesylazide diphosphate for incorporation of prenylazides into a CAAX box-containing peptide using protein farnesyltransferase. J Peptide Res 65:529
Duckworth BP, Xu J, Taton TA, Guo A, Distefano MD (2006) Site-specific, covalent attachment of proteins to a solid surface. Bioconjug Chem 17:967
Duckworth BP, Zhang Z, Hosokawa A, Distefano MD (2007) Selective labeling of proteins by using protein farnesyltransferase. ChemBioChem 8:98
Chan LN, Hart C, Guo L, Nyberg T, Davies BSJ, Fong LG, Young SG, Agnew BJ, Tamanoi F (2009) A novel approach to tag and identify geranylgeranylated proteins. Electrophoresis 30:3598
DeGraw AJ, Palsuledesai C, Ochocki JD, Dozier JK, Lenevich S, Rashidian M, Distefano MD (2010) Evaluation of alkyne-modified isoprenoids as chemical reporters of protein prenylation. Chem Biol Drug Des 76:460
Lin HP, Hsu SC, Wu JC, Sheen IJ, Yan BS, Syu WJ (1999) Localization of isoprenylated antigen of hepatitis delta virus by anti-farnesyl antibodies. J Gen Virol 80:91
Liu X, Suh D, Call J, Prestwich GD (2004) Antigenic prenylated peptide conjugates and polyclonal antibodies to detect protein prenylation. Bioconjug Chem 15:270
Baron R, Fourcade E, Lajoie-Mazenc I, Allal C, Couderc B, Barbaras R, Favre G, Faye JC, Pradines A (2000) RhoB prenylation is driven by the three carboxyl-terminal amino acids of the protein: evidenced in vivo by an anti-farnesyl cysteine antibody. Proc Natl Acad Sci U S A 97:11626
Troutman JM, Roberts MJ, Andres DA, Spielmann HP (2005) Tools to analyze protein farnesylation in cells. Bioconjug Chem 16:1209
Tuinman AA, Thomas DA, Cook KD, Xue CB, Naider F, Becker JM (1991) Mass spectrometric signature of S-prenylated cysteine peptides. Anal Biochem 193:173
Kassai H, Satomi Y, Fukada Y, Takao T (2005) Top-down analysis of protein isoprenylation by electrospray ionization hybrid quadrupole time-of-flight tandem mass spectrometry; the mouse Tgamma protein. Rapid Commun Mass Spectrom 19:269
Appels NMGM, Tung K, Rosing H, Schellens JHM, Beijnen JH (2006) A rapid and simple HPLC-UV method for the determination of inhibition characteristics of farnesyl transferase inhibitors. Biomed Chromatogr 20:161
Appels NMGM, Rosing H, Stephens TC, Hughes A, Schellens JHM, Beijnen JH (2006) Absolute quantification of farnesylated Ras levels in complex samples using liquid chromatography fractionation combined with tryptic digestion and electrospray tandem mass spectrometry. Anal Biochem 352:33
Buser CA, Dinsmore CJ, Fernandes C, Greenberg I, Hamilton K, Mosser SD, Walsh ES, Williams TM, Koblan KS (2001) High-performance liquid chromatography/mass spectrometry characterization of Ki4B-Ras in PSN-1 cells treated with the prenyltransferase inhibitor L-778,123. Anal Biochem 290:126
Nguyen UTT, Guo Z, Delon C, Wu Y, Deraeve C, Fränzel B, Bon RS, Blankenfeldt W, Goody RS, Waldmann H, Wolters D, Alexandrov K (2009) Analysis of the eukaryotic prenylome by isoprenoid affinity tagging. Nat Chem Biol 5:227
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DAV acknowledges research support by the Alfried Krupp von Bohlen und Halbach-Stiftung.
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Nürenberg, G., Volmer, D.A. The analytical determination of isoprenoid intermediates from the mevalonate pathway. Anal Bioanal Chem 402, 671–685 (2012). https://doi.org/10.1007/s00216-011-5262-2
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DOI: https://doi.org/10.1007/s00216-011-5262-2