Abstract
Brief exposure of Chinese hamster ovary cell monolayers prelabeled with [32P]phosphate and [3H]leucine to 1 μg/ml of trypsin under conditions in which cells remain fully viable causes the release of macromolecular 32P and 3H. Whereas ribonuclease treatment was found to affect markedly both the 32P and 3H radioactivity, Pronase treatment had little effect on the macromolecular 32P. Treatment of cells prelabeled with [3H]glucosamine and [32P]phosphate with insolubilized papain also revealed a parallel release of macromolecular glucosamine together with ribonuclease-susceptible macromolecular phosphate. Lactoperoxidase-mediated radioiodination of surface components in cells prelabeled with [32P]phosphate revealed electrophoretic comigration between the 125I and the 32P that are removed from the cells by mild proteolysis. Growth of the cells in Bt2cAMP-testosterone altered the kinetics of release and nature of the macromolecular 32P liberated by proteolysis.
Keywords: putative cell surface RNA, insolubilized papain, dibutyryl cyclic AMP, Chinese hamster ovary cells
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Selected References
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- Bennett M., Mayhew E., Weiss L. RNA in the periphery of rapidly proliferating mouse lymphoid cells. J Cell Physiol. 1969 Oct;74(2):183–190. doi: 10.1002/jcp.1040740210. [DOI] [PubMed] [Google Scholar]
- Carraway K. L., Lam A., Kobylka D., Huggins J. Anomalous staining of membrane lipids on acrylamide gels. Anal Biochem. 1972 Jan;45(1):325–331. doi: 10.1016/0003-2697(72)90036-x. [DOI] [PubMed] [Google Scholar]
- Crooke S. T., Okada S., Busch H. A surface 3'-exonuclease of Novikoff hepatoma ascites cells. Cancer Res. 1972 Aug;32(8):1747–1752. [PubMed] [Google Scholar]
- Davidova S. Y., Shapot V. S. Liporibonucleoprotein complex as an integral part of animal cell plasma membranes. FEBS Lett. 1970 Feb 25;6(4):349–351. doi: 10.1016/0014-5793(70)80096-5. [DOI] [PubMed] [Google Scholar]
- Droege W., Malchow D., Strominger J. L. Cellular heterogeneity in the thymus. I. Electrophoretic analysis of lymphoid cell populations from chickens. Eur J Immunol. 1972 Apr;2(2):156–161. doi: 10.1002/eji.1830020212. [DOI] [PubMed] [Google Scholar]
- 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]
- Glick M. C., Warren L. Membranes of animal cells. 3. Amino acid incorporation by isolated surface membranes. Proc Natl Acad Sci U S A. 1969 Jun;63(2):563–570. doi: 10.1073/pnas.63.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsie A. W., Puck T. T. Morphological transformation of Chinese hamster cells by dibutyryl adenosine cyclic 3':5'-monophosphate and testosterone. Proc Natl Acad Sci U S A. 1971 Feb;68(2):358–361. doi: 10.1073/pnas.68.2.358. [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]
- 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]
- Juliano R., Ciszkowski J., Waite D., Mayhew E. RNA Associated with plasma membranes of ehrlich ascites carcinoma cells. FEBS Lett. 1972 Apr 15;22(1):27–30. doi: 10.1016/0014-5793(72)80210-2. [DOI] [PubMed] [Google Scholar]
- Rieber M., Bacalao J. Alterations in nuclear phosphoproteins of a temperature-sensitive Chinese hamster cell line exposed to non-permissive conditions. Exp Cell Res. 1974 Apr;85(2):334–338. doi: 10.1016/0014-4827(74)90134-7. [DOI] [PubMed] [Google Scholar]
- Rieber M., Bacalao J. Phosphorylation of high-molecular-weight membrane protein species in Chinese-hamster ovary cells in culture: effect of 6-N,2'-O-dibutyryladenosine 3':5'-cyclic monophosphate plus testosterone. Biochem J. 1973 Mar;132(3):641–644. doi: 10.1042/bj1320641. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schneider E. L., Stanbridge E. J., Epstein C. J. Incorporation of 3H-uridine and 3H-uracil into RNA: a simple technique for the detection of mycoplasma contamination of cultured cells. Exp Cell Res. 1974 Mar 15;84(1):311–318. doi: 10.1016/0014-4827(74)90411-x. [DOI] [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]
- Weiss L., Mayhew E. Robonucelic acid within the cellular peripheral zone and the binding of calcium to ionogenic sites. J Cell Physiol. 1967 Jun;69(3):281–291. doi: 10.1002/jcp.1040690304. [DOI] [PubMed] [Google Scholar]
- Zalik S. E., Scott V. Cell surface changes during dedifferentiation in the metaplastic transformation of iris into lens. J Cell Biol. 1972 Oct;55(1):134–146. doi: 10.1083/jcb.55.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
