Skip to main content
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
. 1974 Feb;71(2):489–492. doi: 10.1073/pnas.71.2.489

A Comparison of Membrane Proteins of Normal and Transformed Cells by Lactoperoxidase Labeling

Nancy M Hogg 1
PMCID: PMC388032  PMID: 4360946

Abstract

The enzyme lactoperoxidase (iodide:hydrogen-peroxide oxidoreductase, EC 1.11.1.8) was used to iodinate (125I) accessible proteins on membranes of intact virally transformed and untransformed cells. A number of labeled bands of proteins were detected by acrylamide gel electrophoresis. A heavily labeled band with a molecular weight of approximately 250,000 daltons was found in all untransformed cells but was absent from transformed cells. When Coomassie-blue-stained membrane preparations were compared, a band was seen in normal cells which co-migrated with the lactoperoxidase-labeled band. In transformed cell membranes, three discrete bands were present in the same position. Thus, the expression of this protein may be altered when cells are in the transformed state.

Keywords: iodination, fibroblasts, membranes, gel electrophoresis

Full text

PDF
489

Images in this article

Selected References

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

  1. Barber A. J., Jamieson G. A. Isolation of glycopeptides from low- and high-density platelet plasma membranes. Biochemistry. 1971 Dec 7;10(25):4711–4717. doi: 10.1021/bi00801a018. [DOI] [PubMed] [Google Scholar]
  2. Baur S., Vitetta E. S., Sherr C. J., Schenkein I., Uhr J. W. Isolation of heavy and light chains of immunoglobulin from the surfaces of lymphoid cells. J Immunol. 1971 Apr;106(4):1133–1135. [PubMed] [Google Scholar]
  3. Buck C. A., Glick M. C., Warren L. A comparative study of glycoproteins from the surface of control and Rous sarcoma virus transformed hamster cells. Biochemistry. 1970 Nov 10;9(23):4567–4576. doi: 10.1021/bi00825a016. [DOI] [PubMed] [Google Scholar]
  4. Chiarugi V. P., Urbano P. Electrophoretic analysis of membrane glycoproteins in normal and polyoma virus transformed BHK 21 cells. J Gen Virol. 1972 Feb;14(2):133–140. doi: 10.1099/0022-1317-14-2-133. [DOI] [PubMed] [Google Scholar]
  5. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  6. Glossmann H., Neville D. M., Jr Glycoproteins of cell surfaces. A comparative study of three different cell surfaces of the rat. J Biol Chem. 1971 Oct 25;246(20):6339–6346. [PubMed] [Google Scholar]
  7. Hubbard A. L., Cohn Z. A. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. doi: 10.1083/jcb.55.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Phillips D. R. Effect of trypsin on the exposed polypeptides and glycoproteins in the human platelet membrane. Biochemistry. 1972 Nov 21;11(24):4582–4588. doi: 10.1021/bi00774a025. [DOI] [PubMed] [Google Scholar]
  10. Phillips D. R., Morrison M. The arrangement of proteins in the human erythrocyte membrane. Biochem Biophys Res Commun. 1970 Jul 27;40(2):284–289. doi: 10.1016/0006-291x(70)91007-7. [DOI] [PubMed] [Google Scholar]
  11. Poduslo J. F., Greenberg C. S., Glick M. C. Proteins exposed on the surface of mammalian membranes. Biochemistry. 1972 Jul 4;11(14):2616–2621. doi: 10.1021/bi00764a011. [DOI] [PubMed] [Google Scholar]
  12. Renger H. C., Basilico C. Mutation causing temperature-sensitive expression of cell transformation by a tumor virus (SV40-3T3 mouse cells-growth control). Proc Natl Acad Sci U S A. 1972 Jan;69(1):109–114. doi: 10.1073/pnas.69.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rowe W. P., Pugh W. E., Hartley J. W. Plaque assay techniques for murine leukemia viruses. Virology. 1970 Dec;42(4):1136–1139. doi: 10.1016/0042-6822(70)90362-4. [DOI] [PubMed] [Google Scholar]
  14. Sakiyama H., Burge B. W. Comparative studies of the carbohydrate-containing components of 3T3 and simian virus 40 transformed 3T3 mouse fibroblasts. Biochemistry. 1972 Apr 11;11(8):1366–1377. doi: 10.1021/bi00758a007. [DOI] [PubMed] [Google Scholar]
  15. Schenkein I., Levy M., Uhr J. W. The use of glucose oxidase as a generator of H 2 O 2 in the enzymatic radioiodination of components of cell surfaces. Cell Immunol. 1972 Nov;5(3):490–493. doi: 10.1016/0008-8749(72)90076-7. [DOI] [PubMed] [Google Scholar]
  16. Scher I., Barland P. Studies of surface membranes of two mouse fibroblast cell lines. Biochim Biophys Acta. 1972 Feb 11;255(2):580–588. doi: 10.1016/0005-2736(72)90162-9. [DOI] [PubMed] [Google Scholar]
  17. Shodell M. Environmental stimuli in the progression of BHK-21 cells through the cell cycle. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1455–1459. doi: 10.1073/pnas.69.6.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stanley P., Haslam E. A. The polypeptides of influenza virus. V. Localization of polypeptides in the virion by iodination techniques. Virology. 1971 Dec;46(3):764–773. doi: 10.1016/0042-6822(71)90078-x. [DOI] [PubMed] [Google Scholar]
  19. Warren L., Critchley D., Macpherson I. Surface glycoproteins and glycolipids of chicken embryo cells transformed by a temperature-sensitive mutant of Rous sarcoma virus. Nature. 1972 Feb 4;235(5336):275–278. doi: 10.1038/235275a0. [DOI] [PubMed] [Google Scholar]
  20. Wu H. C., Meezan E., Black P. H., Robbins P. W. Comparative studies on the carbohydrate-containing membrane components of normal and virus-transformed mouse fibroblasts. I. Glucosamine-labeling patterns in 3T3, spontaneously transformed 3T3, and SV-40-transformed 3T3 cells. Biochemistry. 1969 Jun;8(6):2509–2517. doi: 10.1021/bi00834a038. [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