Abstract
Putative cell-surface proteins of tissue-culture cells were identified by lactoperoxidase-catalyzed iodination, a technique that attaches label only to proteins outside the cell membrane. Evidence is presented that these proteins are cell derived, not contaminating serum proteins. On “normal” cells, which exhibit density dependence of growth, one protein of high molecular weight was particularly readily iodinated. This protein was easily removed by mild proteolytic digestion, and, in virus-transformed cells, was either absent or unavailable for iodination. The possible relevance of these observations to the control of growth in cell culture is discussed.
Keywords: fibroblasts, lactoperoxidase, iodination, polyacrylamide gels
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- Arndt-Jovin D. J., Berg P. Quantitative binding of 125 I-concanavalin A to normal and transformed cells. J Virol. 1971 Nov;8(5):716–721. doi: 10.1128/jvi.8.5.716-721.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosmann H. B. Elevated glycosidases and proteolytic enzymes in cells transformed by RNA tumor virus. Biochim Biophys Acta. 1972 Apr 21;264(2):339–343. doi: 10.1016/0304-4165(72)90298-x. [DOI] [PubMed] [Google Scholar]
- Burger M. M. A difference in the architecture of the surface membrane of normal and virally transformed cells. Proc Natl Acad Sci U S A. 1969 Mar;62(3):994–1001. doi: 10.1073/pnas.62.3.994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burger M. M., Bombik B. M., Breckenridge B. M., Sheppard J. R. Growth control and cyclic alterations of cyclic AMP in the cell cycle. Nat New Biol. 1972 Oct 11;239(93):161–163. doi: 10.1038/newbio239161a0. [DOI] [PubMed] [Google Scholar]
- Burger M. M., Goldberg A. R. Identification of a tumor-specific determinant on neoplastic cell surfaces. Proc Natl Acad Sci U S A. 1967 Feb;57(2):359–366. doi: 10.1073/pnas.57.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burger M. M. Proteolytic enzymes initiating cell division and escape from contact inhibition of growth. Nature. 1970 Jul 11;227(5254):170–171. doi: 10.1038/227170a0. [DOI] [PubMed] [Google Scholar]
- Diamond L. Two spontaneously transformed cell lines derived from the same hamster embryo culture. Int J Cancer. 1967 Mar 15;2(2):143–152. doi: 10.1002/ijc.2910020209. [DOI] [PubMed] [Google Scholar]
- 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]
- Inbar M., Sachs L. Interaction of the carbohydrate-binding protein concanavalin A with normal and transformed cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1418–1425. doi: 10.1073/pnas.63.4.1418. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inbar M., Sachs L. Structural difference in sites on the surface membrane of normal and transformed cells. Nature. 1969 Aug 16;223(5207):710–712. doi: 10.1038/223710a0. [DOI] [PubMed] [Google Scholar]
- Klein G. Tumor-specific transplantation antigens: G. H. A. Clowes memorial lecture. Cancer Res. 1968 Apr;28(4):625–635. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- McAllister R. M., Macpherson I. Transformation of a hamster cell line by adenovirus type 12. J Gen Virol. 1968 Jan;2(1):99–106. doi: 10.1099/0022-1317-2-1-99. [DOI] [PubMed] [Google Scholar]
- Nicolson G. L. Topography of membrane concanavalin A sites modified by proteolysis. Nat New Biol. 1972 Oct 18;239(94):193–197. doi: 10.1038/newbio239193a0. [DOI] [PubMed] [Google Scholar]
- Ossowski L., Unkeless J. C., Tobia A., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. II. Mammalian fibroblast cultures transformed by DNA and RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):112–126. doi: 10.1084/jem.137.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ozanne B., Sambrook J. Binding of radioactively labelled concanavalin A and wheat germ agglutinin to normal and virus-transformed cells. Nat New Biol. 1971 Aug 4;232(31):156–160. doi: 10.1038/newbio232156a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Phillips D. R., Morrison M. Exposed protein on the intact human erythrocyte. Biochemistry. 1971 May 11;10(10):1766–1771. doi: 10.1021/bi00786a006. [DOI] [PubMed] [Google Scholar]
- Schnebli H. P. A protease-like activity associated with malignant cells. Schweiz Med Wochenschr. 1972 Aug 19;102(33):1194–1197. [PubMed] [Google Scholar]
- Schnebli H. P., Burger M. M. Selective inhibition of growth of transformed cells by protease inhibitors. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3825–3827. doi: 10.1073/pnas.69.12.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sefton B. M., Rubin H. Release from density dependent growth inhibition by proteolytic enzymes. Nature. 1970 Aug 22;227(5260):843–845. doi: 10.1038/227843a0. [DOI] [PubMed] [Google Scholar]
- Sheppard J. R. Difference in the cyclic adenosine 3',5'-monophosphate levels in normal and transformed cells. Nat New Biol. 1972 Mar 1;236(61):14–16. doi: 10.1038/newbio236014a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
- Unkeless J. C., Tobia A., Ossowski L., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):85–111. doi: 10.1084/jem.137.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warren L., Fuhrer J. P., Buck C. A. Surface glycoproteins of normal and transformed cells: a difference determined by sialic acid and a growth-dependent sialyl transferase. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1838–1842. doi: 10.1073/pnas.69.7.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]