Abstract
The results of metabolic labeling studies and enzymatic treatments followed by analysis on polyacrylamide gels show that the external proteins of hamster fibroblast cell lines, which have been identified by lactoperoxidase-catalyzed iodination, do not contain sulphated mucopolysaccharides or hyaluronic acid and are probably unrelated to collagen. Several of the iodinated species comigrate with carbohydrate-containing molecules. In particular, the major iodine-labeled polypeptide of normal fibroblasts appears to be a glycoprotein. This glycoprotein is absent or much reduced in virus-transformed cells, as detected both by iodination and by metabolic labeling. We conclude that the major iodinated polypeptide is not detected on transformed cells because it is absent rather than because it is masked. Approximate molecular weights of the external proteins are also reported.
Full Text
The Full Text of this article is available as a PDF (734.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bellamy G., Bornstein P. Evidence for procollagen, a biosynthetic precursors of collagen. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1138–1142. doi: 10.1073/pnas.68.6.1138. [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]
- Bretscher M. S. Major human erythrocyte glycoprotein spans the cell membrane. Nat New Biol. 1971 Jun 23;231(25):229–232. doi: 10.1038/newbio231229a0. [DOI] [PubMed] [Google Scholar]
- Bujard E., Mauron J. A two-dimensional separation of acid, neutral and basic amino acids by thin layer chromatography on cellulose. J Chromatogr. 1966 Jan;21(1):19–26. doi: 10.1016/s0021-9673(01)91255-5. [DOI] [PubMed] [Google Scholar]
- Buonassisi V. Sulfated mucopolysaccharide synthesis and secretion in endothelial cell cultures. Exp Cell Res. 1973 Feb;76(2):363–368. doi: 10.1016/0014-4827(73)90388-1. [DOI] [PubMed] [Google Scholar]
- Burger M. M., Martin G. S. Agglutination of cells transformed by Rous sarcoma virus by wheat germ agglutinin and concanavalin A. Nat New Biol. 1972 May 3;237(70):9–12. doi: 10.1038/newbio237009a0. [DOI] [PubMed] [Google Scholar]
- Bussell R. H., Robinson W. S. Membrane proteins of uninfected and Rous sarcoma virus- transformed avian cells. J Virol. 1973 Aug;12(2):320–327. doi: 10.1128/jvi.12.2.320-327.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Dehm P., Jimenez S. A., Olsen B. R., Prockop D. J. A transport form of collagen from embryonic tendon: electron microscopic demonstration of an NH 2 -terminal extension and evidence suggesting the presence of cystine in the molecule (chick embryo-tropocollagen-gel filtration). Proc Natl Acad Sci U S A. 1972 Jan;69(1):60–64. doi: 10.1073/pnas.69.1.60. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furthmayr H., Timpl R. Characterization of collagen peptides by sodium dodecylsulfate-polyacrylamide electrophoresis. Anal Biochem. 1971 Jun;41(2):510–516. doi: 10.1016/0003-2697(71)90173-4. [DOI] [PubMed] [Google Scholar]
- Gahmberg C. G., Hakomori S. I. Altered growth behavior of malignant cells associated with changes in externally labeled glycoprotein and glycolipid. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3329–3333. doi: 10.1073/pnas.70.12.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldberg B., Epstein E. H., Jr, Sherr C. J. Precursors of collagen secreted by cultured human fibroblasts. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3655–3659. doi: 10.1073/pnas.69.12.3655. [DOI] [PMC free article] [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]
- Hughes R. C., Sanford B., Jeanloz R. W. Regeneration of the surface glycoproteins of a transplantable mouse tumor cell after treatment with neuraminidase. Proc Natl Acad Sci U S A. 1972 Apr;69(4):942–945. doi: 10.1073/pnas.69.4.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kraemer P. M. Heparan sulfates of cultured cells. I. Membrane-associated and cell-sap species in Chinese hamster cells. Biochemistry. 1971 Apr 13;10(8):1437–1445. doi: 10.1021/bi00784a026. [DOI] [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]
- Layman D. L., McGoodwin E. B., Martin G. R. The nature of the collagen synthesized by cultured human fibroblasts. Proc Natl Acad Sci U S A. 1971 Feb;68(2):454–458. doi: 10.1073/pnas.68.2.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ossowski L., Quigley J. P., Kellerman G. M., Reich E. Fibrinolysis associated with oncogenic transformation. Requirement of plasminogen for correlated changes in cellular morphology, colony formation in agar, and cell migration. J Exp Med. 1973 Nov 1;138(5):1056–1064. doi: 10.1084/jem.138.5.1056. [DOI] [PMC free article] [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]
- Phillips D. R., Morrison M. Changes in accessibility of plasma membrane protein as the result of tryptic hydrolysis. Nat New Biol. 1973 Apr 18;242(120):213–215. doi: 10.1038/newbio242213a0. [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]
- 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]
- Saito H., Uzman B. G. Production and secretion of chondroitin sulfates and dermatan sulfate by established mammalian cell lines. Biochem Biophys Res Commun. 1971 May 21;43(4):723–728. doi: 10.1016/0006-291x(71)90675-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Schnebli H. P. A protease-like activity associated with malignant cells. Schweiz Med Wochenschr. 1972 Aug 19;102(33):1194–1197. [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]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- Yamagata T., Saito H., Habuchi O., Suzuki S. Purification and properties of bacterial chondroitinases and chondrosulfatases. J Biol Chem. 1968 Apr 10;243(7):1523–1535. [PubMed] [Google Scholar]
- Zavada J., Macpherson I. Transformation of hamster cell lines in vitro by a hamster sarcoma virus. Nature. 1970 Jan 3;225(5227):24–26. doi: 10.1038/225024a0. [DOI] [PubMed] [Google Scholar]