Purification of geranylgeranyl diphosphate synthase from bovine brain

• Autorzy: Sagami H. , Morita Y. , Korenaga T. , Ogura K.
• Języki publikacji: EN

Abstrakty

Geranylgeranyl diphosphate (GGPP) synthase was purified to homogeneity from bovine brain in a one-step affinity column procedure. For the construction of the affinity column, a farnesyl diphosphate (FPP) analog, CM6-amino-l-hexyl)-P- -farnesylmethyl phosphonophosphate, was synthesized and linked to the spacer of the matrix of Affigel 10 via the amino group. The native enzyme appeared to be homooligomer (150-195 kDa) with a molecular mass of the monomer of 37.5 kDa. The pi for the enzyme was 6.2. The Km values for dimethylallyl diphosphate (DMAPP), geranyl diphosphate (GPP) and FTP were estimated to be 33 uM, 0.80 uM and 0.74 uM, respectively. The Km value for isopentenyl diphosphate (IPP) in the presence of both IPP and FPP mixture was 2|iM. The ratio of the reaction velocity for formation of GGPP from DMAPP, GPP or FPP was 0.004:0.145:1. The intermediate FPP was formed in the reaction with GPP as an allylic primer. FPP synthase catalyzing the formation of FPP from DMAPP and IPP was also purified to homogeneity from the same organ by a similar affinity chromatography procedure using a GPP analog, 0-(6-amino-l-hexyl)- -P-geranylmethyl phosphonophosphate as a ligand. The enzyme was a homodimer with a monomeric molecular mass of 40.0 kDa. These results indicate that GGPP, a lipid precursor for the biosynthesis of a majority of prenylated proteins, is synthesized from DMAPP and IPP by the action of FPP synthase catalyzing the reactions C5->C15 followed by the action of GGPP synthase catalyzing the reaction C15->C20.

Słowa kluczowe

EN

bovine brain; geranylgeranyl diphosphate synthase; prenyltransferase; brain

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1994
• Tom: 41
• Numer: 3
• Opis fizyczny: p.293-302,fig.

Twórcy

autor

Sagami H.

• Tohoku University, Katahira, Sendai 980, Japan

autor

Morita Y.

autor

Korenaga T.

autor

Ogura K.

Bibliografia

• 1. Clarke, S. (1992) Protein isoprenylation and methylation at carboxyl-terminal cysteine residues. Awtu. Rev. Biochein. 61,355-386.
• 2. Stimmel, J.B., Deschenes, R.J., Volker, C., Stoch, J. & Clarke, S. (1990) Evidence for an S-farncsyl- cystcine methyl ester at the carboxyl terminus of the Saccharomyces cerevisiae RAS2 protein. Biochemistry 29,9651 -9659.
• 3. Maltese, W.A. (1990) Posttranslational modifi­cation of proteins by isoprenoids in mammalian cells. FASEB J. 4,3319-3327.
• 4. Rine, J. & Kim, S.-H. (1990) A role for isoprenoid lipids in the localization and function of an oncoprotein. New Biol. 2,219-226.
• 5. Takai,Y., Kaibuchi, K., Kikuchi,A. & Kawata, M. (1992) Small GTP-binding proteins. Int. Rev. Cytol. 133,187-230.
• 6. Itoh, T., Kaibuchi, K., Masuda, T., Yamamoto, T., Masuura, Y., Maeda, A., Shimizu, K. & Takai, Y. (1993) The post-translational processing of ras p21 is critical for its stimulation of mitogen-activated protein kinase. /. Biol. Cltern. 268,3025-3028.
• 7. Horiuchi, H., Kaibuchi, K., Kawamura, M., Masuura, Y., Suzuki, N., Kuroda, Y., Kataoka, T. & Takai, Y. (1992) The post-translational processing of ras p21 is critical forits stimulation of yeast adenylate cyclase. Mol. Cell Biol. 12, 4515-4520.
• 8. Kuroda, Y., Suzuki, N. & Kataoka, T. (1993) The effect of posttranslational modifications on the interaction of Ras2 with adenylyl cyclase. Science 259,683-686.
• 9. Boguski, M.S. & McCormick, F. (1993) Proteins regulating Ras and its relatives. Nature (London) 366,643-654.
• 10. Reed, B.C. & Rilling, H.C. (1975) Crystallization and partial characterization of prenyl- transferase from avian liver. Biochemistry 14, 50-54.
• 11. Sagami, H., Ishii, K. & Ogura, K. (1981) Occurrence and unusual properties of geranylgeranyl pyrophosphate synthetase of pig liver. Biochein. Int. 3,669-675.
• 12. Sagami, 11., Korenaga, T., Ogura, K., Stcigcr, A., Pyun, H.-J. & Coates, R.M. (1992) Studies on geranylgeranyl diphosphate synthase from rat liver: specific inhibition by 3-azageranylgeranyl diphosphate. Arch. Biochem. Biophys. 297, 314-320.
• 13. Sagami, H., Korenaga, T. & Ogura, K. (1993) Geranylgeranyl diphosphate synthase cata­lyzing the single condensation between isopentenyl diphosphate and famesyl diphos­phate. /. Biochein. 114,118-121.
• 14. Rilling, H.C., Bruenger, E., Epstein, W.VV. & Crain, P.F. (1990) Prenylated proteins: structure of the isoprenoid group. Science 247, 31&-320.
• 15. Famsworth, C.C., Gelb, M.H. & Glomset, J.A. (1990) Identification of geranylgeranyl- -modified proteins in HeLa cells. Science 247, 320-322.
• 16. Epstein, W.W., Lever, D., Leining, L.M., Bruenger, E. & Rilling, H.C. (1991) Quantitation of prenylcysteines by a selective cleavage reaction. Proc. Natl. Acad. Sci. U.S.A. 88, 9668-9670.
• 17. Davisson, V.J., Woodside, A.B. & Poulter, C.D. (19S5) Synthesis of allylic and homoallylic isoprenoid pyrophoshate. Methods Enzymol. 110, 130-144.
• 18. Bartlett, D.L., King, C.-H.R. & Poulter, C.D. (1985) Purification of farnesyl pyrophosphate synthetase by affinity chromatography. Methods Enzymol. 110,171-184.
• 19. Fujii, H., Koyama, T. & Ogura, K. (1982) Efficient hydrolysis of poly prenyl pyrophosphates. Biochim. Biophys. Acta 712,716-718.
• 20. Dogbo, O. & Camara, B. (1987) Purification of isopentenyl pyrophosphate isomerase and geranylgeranyl pyrophosphate synthase from Capsicum chromoplasts by affinity chroma­tography. Biochim. Biophys. Acta 920,140-148.
• 21. Laferrifcre, A. & Beyer, P. (1991) Purification of geranylgeranyl diphosphate synthase from Sinapis alba etioplasts. Biochim. Biophys. Acta 1077,167-172.
• 22. Brinkhaus, F.L. & Rilling, H.C. (1988) Purifica­tion of geranylgeranyl diphosphate synthase from Phycomyces blakesleanus. Arch. Biochem. Biophys. 266, 607-612.
• 23. Chen, A. & Poulter, C.D. (1993) Purification and characterization of farnesyl diphosphate/ge- ranylgeranyl diphosphate synthase. J. Biol. Chem. 268,11002-11007.
• 24. Tachibana, A., Tanaka, T., Taniguchi, M. & Oi, S. (1993) Purification and characterization of geranylgeranyl diphosphate synthase from Methanobacterium thennoformicicum SF-4. Biosci. Biotech. Biochem. 57,1129-1133.