Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 May;73(5):1523–1527. doi: 10.1073/pnas.73.5.1523

High in vivo rates of methionine biosynthesis in transformed human and malignant rat cells auxotrophic for methionine.

R M Hoffman, R W Erbe
PMCID: PMC430329  PMID: 179090

Abstract

Unlike normal cells, malignant rat and two simian virus 40-transformed human cell lines can neither grow nor survive in B12-and folate-supplemented media in which methionine is replaced by homocysteine. Yet three lines of evidence indicate that the malignant and transformed cells synthesize large amounts of methionine endogenously through the reaction catalyzed by 5-methyltetrahydropteroyl-L-glutamate; L-homocysteine S-methyltransferase (EC 2.1.1.13). (1) The activities of this methyltransferase were comparable in extracts of malignant and normal cells. (2) The uptake of radioactive label from [5-14C]methyltetrahydropteroyl-L-glutamic acid (5-Me-H4PteGlu) was at least as great in the malignant cells as in the normals and was nearly totally dependent on the addition of homocysteine, the methyl acceptor; furthermore, 59-84% of the label incorporated by cells was recovered as methionine.

Full text

PDF
1523

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashe H., Clark B. R., Chu F., Hardy D. N., Halpern B. C., Halpern R. M., Smith R. A. N5-methyltetrahydrofolate: homocysteine methyltransferase activity in extracts from normal, malignant and embryonic tissue culture cells. Biochem Biophys Res Commun. 1974 Mar 25;57(2):417–425. doi: 10.1016/0006-291x(74)90947-4. [DOI] [PubMed] [Google Scholar]
  2. Chello P. L., Bertino J. R. Dependence of 5-methyltetrahydrofolate utilization by L5178Y murine leukemia cells in vitro on the presence of hydroxycobalamin and transcobalamin II. Cancer Res. 1973 Aug;33(8):1898–1904. [PubMed] [Google Scholar]
  3. EAGLE H., PIEZ K. The population-dependent requirement by cultured mammalian cells for metabolites which they can synthesize. J Exp Med. 1962 Jul 1;116:29–43. doi: 10.1084/jem.116.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Erbe R. W. Inborn errors of folate metabolism (second of two parts). N Engl J Med. 1975 Oct 16;293(16):807–812. doi: 10.1056/NEJM197510162931606. [DOI] [PubMed] [Google Scholar]
  5. GIRARDI A. J., JENSEN F. C., KOPROWSKI H. SV40-INDUCED TRANFORMATION OF HUMAN DIPLOID CELLS: CRISIS AND RECOVERY. J Cell Physiol. 1965 Feb;65:69–83. doi: 10.1002/jcp.1030650110. [DOI] [PubMed] [Google Scholar]
  6. Halpern B. C., Clark B. R., Hardy D. N., Halpern R. M., Smith R. A. The effect of replacement of methionine by homocystine on survival of malignant and normal adult mammalian cells in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1133–1136. doi: 10.1073/pnas.71.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Halpern B. C., Ezzell R., Hardy D. N., Clark B. R., Ashe H., Halpern R. M., Smith R. A. Effect of methionine replacement by homocystine in cultures containing both malignant rat breast carcinosarcoma (Walker-256) cells and normal adult rat liver fibroblasts. In Vitro. 1975 Jan-Feb;11(1):14–19. doi: 10.1007/BF02615317. [DOI] [PubMed] [Google Scholar]
  8. Halpern R. M., Halpern B. C., Clark B. R., Ashe H., Hardy D. N., Jenkinson P. Y., Chou S. C., Smith R. A. New approach to antifolate treatment of certain cancers as demonstrated in tissue culture. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4018–4022. doi: 10.1073/pnas.72.10.4018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kamely D., Littlefield J. W., Erbe R. W. Regulation of 5-methyltetrahydrofolate: homocysteine methyltransferase activity by methionine, vitamin B12, and folate in cultured baby hamster kidney cells. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2585–2589. doi: 10.1073/pnas.70.9.2585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Rosenblatt D. S., Erbe R. W. Reciprocal changes in the levels of functionally related folate enzymes during the culture cycle in human fibroblasts. Biochem Biophys Res Commun. 1973 Oct 15;54(4):1627–1633. doi: 10.1016/0006-291x(73)91173-x. [DOI] [PubMed] [Google Scholar]
  12. Taylor R. T., Hanna M. L. 5-Methyltetrahydrofolate aromatic alkylamine N-methyltransferase: an artefact of 5,10-methylenetetrahydrofolate reductase activity. Life Sci. 1975 Jul 1;17(1):111–119. doi: 10.1016/0024-3205(75)90246-5. [DOI] [PubMed] [Google Scholar]
  13. Todaro G. J., Meyer C. A. Transformation assay for murine sarcoma viruses using a Simian virus 40-transformed human cell line. J Natl Cancer Inst. 1974 Jan;52(1):167–171. doi: 10.1093/jnci/52.1.167. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES