Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Inhibition of C-terminal O-methyltransferase by a rat liver cytosolic peptide

Abstract

The activity of S-famesylcysteine O-methyltransferase was assayed by incubating the enzyme with a syntheticin vitro substrate, [N-acetyl-S-trans, trans-famesyl-L-cysteine (AFC)], together with S-adenosyl-L-[methyl-14C]methionine. The resulting methylesterification product, [N-acetyl-S-trans, trans-famesyl-L-cysteine (methyl-14C) ester (AFCME)], was then analyzed either directly on HPLC or by converting the AFC[methyl-14C]ME to [methyl-14C] alcohol by basehydrolysis. Employing these two analytical methods, it was established that a peptide purified from rat liver cytosol fraction [Int. J. Biochem., 25, 1157 (1993)] strongly inhibited the above enzyme activity with IC50 of 7.1X10−8 M. Also, the S-famesylcysteine O-methyltransferase from several human colon cancer cells was also equally inhibited by the peptide.

This is a preview of subscription content, log in to check access.

References Cited

  1. Barbacid, M., ras GENES.Ann. Rev. Biochem., 56, 779–828 (1987).

  2. Bos, J. L., Fearon, E. R. and Hamilton, S. R., Prevalence of ras gene mutation in human colorectal cancer.Nature, 327, 293–297 (1987).

  3. 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).

  4. Chiva, V. A. and Mato, J. M., Inhibition of phospholipid methylation by a cytosolic factor.Biochem. J., 218, 637–639 (1984).

  5. Clarke, S., Protein isoprenylation and methylation at carboxyl-terminal cysteine residues.Ann. Rev. Biochem., 61, 355–386 (1992).

  6. Gibbs, J. B., Ras C-terminal processing enzymes; New drug targets?Cell, 65, 1–4 (1991).

  7. Haubruck, H. and McCormick, F., Ras p21; effects and regulation.Biochim. Biophys. Acta, 1072, 215–229 (1991).

  8. Hong, S. Y., Lee, H. W., Desi, S., Kim, S. and Paik, W. K., Studies on naturally occuring proteinous inhibitor for transmethylation reactions.Eur. J. Biochem., 156, 79–84 (1986).

  9. Kim, S. and Paik, W. K., Purification and assay of Protein methylase II(S-Adenosylmethionine: protein-car-boxylmethyltransferase; EC 2.1.1.24). In Stein G., Stein J. and Kleinsmith L. (Eds),Methods in Cell Biology, Vol. 19, Academic Press, New York 1978, pp. 79–88.

  10. Kim, S., S-Adenosylmethionine: protein-carboxyl O-methyltransferase (Protein Methylase II). In Wold F. and Moldave K. (Eds),Methods in Enzymology, Vol. 106, Academic Press, New York, 1984, pp. 295–309.

  11. Lowy, D. R. and Willumsen, B. M., Function and regulation of RAS.Ann. Rev. Biochem., 62, 851–891 (1993).

  12. Rodenhuis, S. and Slebos, R. J. C., The ras oncogenes in human lung cancer.Am. Rev. Respir. Dis., 142, S27-S30 (1990).

  13. Park, S. H., Lee, H. W., Kim, S. and Paik, W. K., Peptide inhibitor for S-adenosyl-L-methionine dependent transmethylation reactions.Int. J. Biochem., 25, 1157–1164 (1993).

  14. Stephenson, R. C. and Clarke, S., Identification of a C-terminal protein carboxyl methyltransferase in rat liver membranes utilizing a synthetic famesylcysteine containing peptide substrate.J. Biol. Chem., 265, 16248–16254 (1990).

  15. Volker, C., Miller, R. A. and Stock, J. B., S-Famesylcysteine methyltransferase in the bovine brain.Methods: A Companion to Methods in Enzymology, 1, 283–287 (1990).

Download references

Author information

Correspondence to Seung Hee Park or Hyang Woo Lee.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Park, S.H., Lee, H.W. Inhibition of C-terminal O-methyltransferase by a rat liver cytosolic peptide. Arch. Pharm. Res. 17, 354 (1994). https://doi.org/10.1007/BF02974176

Download citation

Key words

  • Peptide inhibitor
  • S-famesylcysteine O-methyltransferase