Archives of Pharmacal Research

The Pharmaceutical Society of Korea (대한약학회)
권호 표지

Protein Carboxylmethylation in Porcine Spleen is Mainly Mediated by Class I Protein Carboxyl O-Methyltransferase

  • Cho, Jae-Youl (Department of Genetic Engineering, Faculty of Life Science and Technology, Sungkyunkwan University, School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Kim, Sung-Soo (Department of Anatomy, Ajou University School of Medicine) ;
  • Kwon, Myung-Hee (Department of Microbiology, Ajou University School of Medicine) ;
  • Kim, Seong-Hwan (Department of Genetic Engineering, Faculty of Life Science and Technology, Sungkyunkwan University) ;
  • Lee, Hyang-Woo (Department of Biochemistry and Molecular Biology, College of Pharmacy, Sungkyunkwan University) ;
  • Hong, Sung-Youl (Department of Genetic Engineering, Faculty of Life Science and Technology, Sungkyunkwan University)
  • Published : 2004.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

The functional role of protein carboxylmethylation (PCM) has not yet been clearly elucidated in the tissue level. The biochemical feature of PCM in porcine spleen was therefore studied by investigating the methyl accepting capacity (MAC) of natural endogenous substrate proteins for protein carboxyl O-methyltransferase (PCMT) in various conditions. Strong acidic and alkaline-conditioned (at pH 11.0) analyses of the MAC indicated that approximately 65％ of total protein methylation seemed to be mediated by spleen PCMT. The hydrolytic kinetics of the PCM products, such as carboxylmethylesters (CMEs), under mild alkaline conditions revealed that there may be three different kinds of CMEs [displaying half-times (T$_{1}$2/) of 1.1 min (82.7％ of total CMEs), 13.9 min (4.6％), and 478.0 min (12.7％)], assuming that the majority of CME is base-labile and may be catalyzed by class I PCMT. In agreement with these results, several natural endogenous substrate proteins (14, 31 and 86 kDa) were identified strikingly by acidic-conditioned electrophoresis, and their MAC was lost upon alkaline conditions. On the other hand, other proteins (23 and 62 kDa) weakly appeared under alkaline conditions, indicating that PCM mediated by class II or III PCMT may be a minor reaction. The MAC of an isolated endogenous substrate protein (23-kDa) was also detected upon acidic-conditioned electrophoresis. Therefore, our date suggest that most spleen PCM may be catalyzed by class I PCMT, which participates in repairing aged proteins.

Keywords

References

  1. Aswad, D. W., Protein carboxyl methylation in eukaryotes. Curr. Opin. Cell Biol., 1, 1182-1187 (1989) https://doi.org/10.1016/S0955-0674(89)80069-9
  2. Aswad, D. W. and Deight, E. A., Endogenous substrates for protein carboxyl methyltransferase in cytosolic fractions of bovine brain. J. Neurochem., 41, 1702-1709 (1983) https://doi.org/10.1111/j.1471-4159.1983.tb00883.x
  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., 42, 248-254 (1976)
  4. Clarke S., Aging as war between chemical and biochemical processes: Protein methylation and the recognition of age-damaged proteins for repair. Ageing Res. Rev., 2, 263-285 (2003) https://doi.org/10.1016/S1568-1637(03)00011-4
  5. Cho, J. Y., Kim, S., Lee, H. W. and HongS., Protein carboxyl O-methylation in porcine liver and testis. Yakhak Hoeji, 45, 46-54 (2001)
  6. Diliberto, E. J., JR, Viveros, O. H., and Axelrod, J., Subcellular distribution of protein carboxymethylase and its endogenous substrates in the adrenal medulla: Possible role in exitation-secretion coupling. Proc. Natl. Acad. Sci. U.S.A., 73, 4050-4054 (1976) https://doi.org/10.1073/pnas.73.11.4050
  7. Diliberto, E. J. and Axelrod, J., Regional and subcellular distribution of protein carboxymethylase in brain and other tissues. J. Neurochem., 26, 1159-1165 (1976) https://doi.org/10.1111/j.1471-4159.1976.tb07001.x
  8. Fairbanks, G. and Avruch, J., Four gel systems for elec-trophoretic fractionation of membrane proteins using ionic detergents. J. Supramol. Struct., 1, 66-75 (1972) https://doi.org/10.1002/jss.400010110
  9. Farrar, C. and Clarke, S., Altered levels of S-adenosyl-methionine and S-adenosylhomocysteine in the brains of L-isoaspartyl (D-aspartyl) O-methyltransferase-deficient mice. J. Biol. Chem., 277, 27856-27863 (2002) https://doi.org/10.1074/jbc.M203911200
  10. Gagnon, C., Sherins, R. J., Philips, D. M., and Bardin, C. W., Deficiency of protein-carboxyl methylase in immotile spermatozoa of infertile men. N. Engl. J. Med., 306, 821-825 (1982) https://doi.org/10.1056/NEJM198204083061401
  11. Gingras, D., Menard, P., and Beliveau, R., Protein carboxyl methylation in kidney brush-border membranes. Biochem. Biophy Acta, 1066, 261-277 (1991) https://doi.org/10.1016/0005-2736(91)90196-F
  12. Hong, S. Y., Lee, H. W., Desi, S., Kim, S., and Paik, W. K., Studies on naturally occurring proteinous inhibitor for transmethylation reactions. Eur. J. Biochem., 156, 79-84 (1986) https://doi.org/10.1111/j.1432-1033.1986.tb09551.x
  13. Hrycyna, C. A. and Clarke, S., Modification of eukaryotic signaling protein by C-terminal methylation reactions. Pharmac. Ther., 59, 281-300 (1993) https://doi.org/10.1016/0163-7258(93)90071-K
  14. Ingrosso, D., D'angelo, S., di Carlo, E., Perna, A. F., Zappia, V., and Galletti, P., Increased methyl esterification of altered aspartyl residues in erythrocyte membrane proteins in response to oxidative stress. Eur. J. Biochem., 267, 4397-405 (2000) https://doi.org/10.1046/j.1432-1327.2000.01485.x
  15. Janson, C. A. and Clarke, S., Identification of aspartic acid as a site of methylation in human erythrocyte membrane protein. J. Biol. Chem., 255, 11640-11643 (1980)
  16. Kim, S., Cho, J., Lee, H. W., and Hong, S., Purification and properties of protein methylase II from porcine spleen. Kor. J. Biochem.,27, 179-184 (1994)
  17. Kim, S., Nochumson, S., Chin, W., and Paik, W. K., A rapid method for the purification S-adenosyl-L-methionine: Protein carboxyl O-methyltransferase by affinity chromatography. Anal. Biochem., 84, 415-422 (1978) https://doi.org/10.1016/0003-2697(78)90059-3
  18. Kowluru, A., Seavey, S. E., Li, G., Sorenson, R. L., Weinhaus, A. J., Nesher, R., Rabaglia, M. E., Vadakekalam, J., and Metz, S. A., Glucose- and GTP-dependent stimulation of the carboxyl methylation of CDC42 in rodent and human pancreatic islets and pure beta cells. Evidence for an essential role of GTP-binding proteins in nutrient-induced insulin secretion. J. Clin. Invest., 98, 540-555 (1996) https://doi.org/10.1172/JCI118822
  19. Kowluru, A. and Amin, R., Inhibitors of post-translational modifications of G-proteins as probes to study the pancreatic beta cell function: potential therapeutic implications. Curr. Drug Targets Immune. Endocr. Metabol. Disord., 2, 129-139 (2002) https://doi.org/10.2174/1568008023340668
  20. Krebs, E. G. and Beavo, J. A., Phosphorylation-dephosphory-lation of enzymes. Annu. Rev. Biochem., 48, 923-959 (1979) https://doi.org/10.1146/annurev.bi.48.070179.004423
  21. Kwon, M., Jung, K, Lee, H. Y., Lee, H. W., and Hong, S., Purification and characterization of protein methylase II inhibitor from procine liver. Korean Biochem. J., 27, 569-575 (1994)
  22. Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-683 (1970) https://doi.org/10.1038/227680a0
  23. Leonard, E. J., Skeel, A., Chiang, P. K., and Cantoni, G. L., The action of the adenosylhomocysteine hydrolase inhibitory 3-deazaadenosine, on phagocytic function of mouse macrophages and human monocytes. Biochem. Biophys. Res. Commun., 84, 102-109 (1978) https://doi.org/10.1016/0006-291X(78)90269-3
  24. Law, R. E., Stimmel, J. B., Damore, M. A., Carter, C., Clarke, S., and Wall, R., Lippolysaccharide-induced NF-kB activation in mouse 70Z/3 pre-B lymphocytes is inhibited by mevinolin and 5'-methyl thioadenosine: Role of protein isoprenylation and carboxyl methylation reaction. Mol. Cell Biol., 12, 103-111 (1992)
  25. Lee, J. and Stock, J., Protein phosphatase 2A catalytic subunit is methyl-esterified at its carboxyl terminus by a novel methyltransferase. J. Biol. Chem., 268, 19192-19195 (1993)
  26. Najbauer, J., Johnson, B. A., and Aswad, D.W., Amplification and detection of substrates for protein carboxyl methyltrans-ferase in PC12 cells. Anal. Biochem., 197, 412-420 (1991) https://doi.org/10.1016/0003-2697(91)90413-N
  27. O'Connor, C. M. and Clarke, S., Carboxyl methylation of cytosolic proteins in intact human erythrocytes. J. Biol. Chem., 259, 2570-2578 (1984)
  28. O'Dea, R. F., Viveros, O. H., and Diliberto, E. J., Jr., Protein carboxyl methylation: Role in the regulation of cell functions. Biochem. Pharmacol., 30, 1163-1168 (1981) https://doi.org/10.1016/0006-2952(81)90292-6
  29. Paik, W. K. and Kim, S., Reevaluation of enzymology of protein methylation. In: Protein Methylation. Paik, W. K. and Kim, S. (Eds) CRC Press, Boca Raton, Florida, pp.1-2 (1990)
  30. Paik, W. K. and Kim S., Effect of methyl substitution on protein tertiary structure. J. Theor. Biol., 155,335-342 (1992) https://doi.org/10.1016/S0022-5193(05)80602-2
  31. Park, S., Lee, H. W., Kim, S., and Paik, W. K., A peptide inhibitor for S-adenosyl-L-methionine-dependent transmethylation reactions. lnt. J. Biochem., 1157-1164 (1993) https://doi.org/10.1016/0020-711X(93)90594-5
  32. Pike, M. C., Kredich, N. M., and Synderman, R, Requirement of S-adenosyl-L-methionine-mediated methylation for human monocyte chemotaxis. Proc. Natl. Acad. Sci. U.S.A., 75, 3928-3932 (1978) https://doi.org/10.1073/pnas.75.8.3928
  33. Rao, A. and Reithmeier, R. A., Reactive sulfhydryl groups of the band 3 polypeptide from human erythroycte membranes. Location in the primary structure. J. Biol. Chem., 254, 6144-6150 (1979)
  34. Rodriguez, A. B., Barriga, C., and De la Fuente, M., Mechanisms of action involved in the chemoattractant activity of three beta-Iactamic antibiotics upon human neutrophils. Biochem. Pharmacol., 41, 931-936 (1991) https://doi.org/10.1016/0006-2952(91)90198-E
  35. Seo, D. W., Kim, Y. K., Cho, E. J., Han, J. W., Lee, H. Y., Hong, S., and Lee, H. W., Oligosaccharide-linked acyl carrier protein, a novel transmethylase inhibitor, from porcine liver inhibits cell growth. Arch. Pharm. Res., 25, 463-468 (2002) https://doi.org/10.1007/BF02976603
  36. Terwilliger, T. C. and Clarke, S., Methylation of membrane protein in human erythrocytes. J. Biol. Chem., 257, 3067-3076 (1981)
  37. Veeraragavan, K. and Gagnon, C., Mammalian protein methylesterase, Biochem. J., 260, 11-17 (1989)
  38. Vafai, S. B. and Stock, J. B., Protein phosphatase 2A methylation: a link between elevated plasma homocysteine and Alzheimer's Disease. FEBS Lett., 518, 1-4 (2002) https://doi.org/10.1016/S0014-5793(02)02702-3
  39. Vincent, P. L. and Siegel, F. L., Carboxylmethylation of calmodulin in cultured pituitary cells. J. Neurochem., 49, 1613-1622 (1987) https://doi.org/10.1111/j.1471-4159.1987.tb01035.x
  40. Winter-Vann, A. M., Kamen, B. A., Bergo, M. O., Young, S. G., Melnyk, S., James, S. J., and Casey, P. J., Targeting Ras signaling through inhibition of carboxyl methylation: an unexpected property of methotrexate. Proc. Natl. Acad. Sci. U. S. A., 100, 6529-6534 (2003) https://doi.org/10.1073/pnas.1135239100
  41. Xie, H. and Clarke, S., Protein phosphatase 2A is reversibly modified by methyl esterification at its C-terminal leucine residue in bovine brain. J. Biol. Chem., 269, 1981-1984 (1994)
  42. Yamane, H. K., Farnsworth, C. C., Xie, H., Evans, T., Howald, W. N., Gelb, M. H., Glomset, J. A., Clarke, S., and Fung, B. K. K., Membrane binding domain of the small G protein G25K contains an S-(all-trans-geranylgeranyl)cysteine methyl ester at its carboxyl terminus. Proc. Natl. Acad. Sci. U.S.A., 88, 286-290 (1991) https://doi.org/10.1073/pnas.88.1.286
  43. Zolnierowicz, S., Type 2A protein phosphatase, the complex regulator of numerous signaling pathways. Biochem. Pharmacal., 60,1225-1235 (2000) https://doi.org/10.1016/S0006-2952(00)00424-X