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. 1980 Feb;33(2):597–605. doi: 10.1128/jvi.33.2.597-605.1980

Envelope polypeptides of Friend leukemia virus: purification and structural analysis.

J Schneider, H Falk, G Hunsmann
PMCID: PMC288584  PMID: 7411688

Abstract

Roughly 10% of surface glycoproteins in the envelope of mature Friend murine leukemia virus are coupled to membrane polypeptides by disulfide bridges. The remaining 90% of these glycoproteins are associated noncovalently. However, they could also be linked to membrane polypeptides by the treatment of purified Friend murine leukemia virus with 2,2'dithiobis(m-nitropyridine). These amphiphilic heterodimer polypeptides, gp84/86, were recovered almost quantitatively in the form of aggregates, termed rosettes, when prepared by solubilization of the viral membrane with Triton X-100 and subsequent velocity sedimentation. gp69/71 and p12(E)/15(E) were purified from these protein micelles after reduction of the disulfide bonds by gel chromatography. Electron micrographs of rosettes, as well as of purified p12(E)/15(E), showed structures different from native viral knobs. Isolated gp84/86 could be reassociated and then displayed more similarity to these viral surface projections. As shown by peptide mapping, the primary structures of the glycoproteins gp69/71 are highly related as are those of the membrane polypeptides p12(E) and p15(E). Furthermore, it was shown by two-dimensional polyacrylamide gel electrophoresis and re-electrophoresis of purified gp84/86 that the larger component, gp86, was composed of gp71 associated with p15(E) and p12(E), whereas the smaller component, gp84, was formed by gp69 bound only to p12(E).

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Selected References

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  1. Bartholomew R. M., Esser A. F., Müller-Eberhard H. J. Lysis of oncornaviruses by human serum. Isolation of the viral complement (C1) receptor and identification as p15E. J Exp Med. 1978 Mar 1;147(3):844–853. doi: 10.1084/jem.147.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Larco J. E., Rapp U. R., Todaro G. J. Cell surface receptors for ecotropic MuLV: detection and tissue distributions of free receptors in vivo. Int J Cancer. 1978 Mar 15;21(3):356–360. doi: 10.1002/ijc.2910210317. [DOI] [PubMed] [Google Scholar]
  4. Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
  5. Famulari N. G., Buchhagen D. L., Klenk H. D., Fleissner E. Presence of murine leukemia virus envelope proteins gp70 and p15(E) in a common polyprotein of infected cells. J Virol. 1976 Nov;20(2):501–508. doi: 10.1128/jvi.20.2.501-508.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frank H., Schwarz H., Graf T., Schäfer W. Properties of mouse leukemia viruses. XV. Electron microscopic studies on the organization of Friend leukemia virus and other mammalian C-type viruses. Z Naturforsch C. 1978 Jan-Feb;33(1-2):124–138. doi: 10.1515/znc-1978-1-224. [DOI] [PubMed] [Google Scholar]
  7. Helenius A., von Bonsdorff C. H. Semlike Forest virus membrane proteins. Preparation and characterization of spike complexes soluble in detergent-free medium. Biochim Biophys Acta. 1976 Jul 15;436(4):895–899. doi: 10.1016/0005-2736(76)90421-1. [DOI] [PubMed] [Google Scholar]
  8. Hunsmann G., Moennig V., Pister L., Seifert E., Schäfer W. Properties of mouse leukemia viruses. VIII. The major viral glycoprotein of Friend leukemia virus. Seroimmunological, interfering and hemagglutinating capacities. Virology. 1974 Dec;62(2):307–318. doi: 10.1016/0042-6822(74)90394-8. [DOI] [PubMed] [Google Scholar]
  9. Ihle J. N., Denny T. P., Bolognesi D. P. Purification and serological characterization of the major envelope glycoprotein from AKR murine leukemia virus and its reactivity with autogenous immune sera from mice. J Virol. 1976 Mar;17(3):727–736. doi: 10.1128/jvi.17.3.727-736.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ihle J. N., Hanna M. G., Jr Natural immunity to endogenous oncornaviruses in mice. Contemp Top Immunobiol. 1977;6:169–174. doi: 10.1007/978-1-4684-3051-6_5. [DOI] [PubMed] [Google Scholar]
  11. Ihle J. N., Jr, Hanna M. G., Schafer W., Hunsmann G., Bolognesi D. P., HUPER G. Polypeptides of mammalian oncornaviruses. III. Localization of p 15 and reactivity with natural antibody. Virology. 1975 Jan;63(1):60–67. doi: 10.1016/0042-6822(75)90370-0. [DOI] [PubMed] [Google Scholar]
  12. Ikeda H., Hardy W., Jr, Tress E., Fleissner E. Chromatographic separation and antigenic analysis of proteins of the oncornaviruses. V. Identification of a new murine viral protein, p15(E). J Virol. 1975 Jul;16(1):53–61. doi: 10.1128/jvi.16.1.53-61.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Karshin W. L., Arcement L. J., Naso R. B., Arlinghaus R. B. Common precursor for Rauscher leukemia virus gp69/71, p15(E), and p12(E). J Virol. 1977 Sep;23(3):787–798. doi: 10.1128/jvi.23.3.787-798.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kende M., Hill R., Dinowitz M., Stephenson J. R., Kelloff G. J. Naturally occurring lymphocyte-mediated immunity to endogenous type-C virus in the mouse. Blocking of the lymphocyte reactivity with antisera to the virus. J Exp Med. 1979 Feb 1;149(2):358–371. doi: 10.1084/jem.149.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kennel S. J. Purification of a glycoprotein from mouse ascites fluid by immunoaffinity chromatography which is related to the major glycoprotein of murine leukemia viruses. Immunologic and structural comparison with purified viral glycoproteins. J Biol Chem. 1976 Oct 25;251(20):6197–6204. [PubMed] [Google Scholar]
  16. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  17. Krantz M. J., Strand M., August J. T. Biochemical and immunological characterization of the major envelope glycoprotein gp69/71 and degradation fragments from Rauscher leukemia virus. J Virol. 1977 Jun;22(3):804–815. doi: 10.1128/jvi.22.3.804-815.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Leamnson R. N., Shander M. H., Halpern M. S. A structural protein complex in Moloney leukemia virus. Virology. 1977 Jan;76(1):437–439. doi: 10.1016/0042-6822(77)90318-x. [DOI] [PubMed] [Google Scholar]
  20. Moennig V., Frank H., Hunsmann G., Schneider I., Schafer W. Properties of mouse leukemia viruses. VII. The major viral glycoprotein of friend leukemia virus. Isolation and physicochemical properties. Virology. 1974 Sep;61(1):100–111. doi: 10.1016/0042-6822(74)90245-1. [DOI] [PubMed] [Google Scholar]
  21. Moennig V., Hunsmann G., Schäfer W. Partielle Reinigung und biologisch-serologische Charakterisierung Kohlenhydrat-haltiger Komponenten aus Präparaten von Friend-Leukämie-Virus. Z Naturforsch C. 1973 Nov-Dec;28(11):785–788. [PubMed] [Google Scholar]
  22. Montelaro R. C., Sullivan S. J., Bolognesi D. P. An analysis of type-C retrovirus polypeptides and their associations in the virion. Virology. 1978 Jan;84(1):19–31. doi: 10.1016/0042-6822(78)90215-5. [DOI] [PubMed] [Google Scholar]
  23. Murray M. J., Kabat D. Genetic and sialylation sources of heterogeneity of the murine leukemia virus membrane envelope glycoproteins gp69/71. J Biol Chem. 1979 Feb 25;254(4):1340–1348. [PubMed] [Google Scholar]
  24. Naso R. B., Arcement L. J., Karshin W. L., Jamjoom G. A., Arlinghaus R. B. A fucose-deficient glycoprotein precursor to Rauscher leukemia virus gp69/71. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2326–2330. doi: 10.1073/pnas.73.7.2326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pinter A., Fleissner E. Structural studies of retroviruses: characterization of oligomeric complexes of murine and feline leukemia virus envelope and core components formed upon cross-linking. J Virol. 1979 Apr;30(1):157–165. doi: 10.1128/jvi.30.1.157-165.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pinter A., Fleissner E. The presence of disulfide-linked gp70-p15(E) complexes in AKR murine leukemia virus. Virology. 1977 Dec;83(2):417–422. doi: 10.1016/0042-6822(77)90187-8. [DOI] [PubMed] [Google Scholar]
  27. Pinter A., Lieman-Hurwitz J., Fleissner E. The nature of the association between the murine leukemia virus envelope proteins. Virology. 1978 Dec;91(2):345–351. doi: 10.1016/0042-6822(78)90382-3. [DOI] [PubMed] [Google Scholar]
  28. Schafer W., Hunsmann G., Moennig V., Noranha F., Bolognesi D. P., Green R. W., Hüper G. Polypeptides of mammalian oncornaviruses. II Characterization of murine leukemia virus polypeptide (p 15) bearing interspecies reactivity. Virology. 1975 Jan;63(1):48–59. doi: 10.1016/0042-6822(75)90369-4. [DOI] [PubMed] [Google Scholar]
  29. Schneider J., Hunsmann G. Surface expression of murine leukemia virus structural polypeptides on host cells and the virion. Int J Cancer. 1978 Aug 15;22(2):204–213. doi: 10.1002/ijc.2910220215. [DOI] [PubMed] [Google Scholar]
  30. Schneider J., Schwarz H., Hunsmann G. Rosettes from Friend leukemia virus envelope: preparation and physicochemical and partial biological characterization. J Virol. 1979 Feb;29(2):624–632. doi: 10.1128/jvi.29.2.624-632.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shapiro S. Z., Strand M., August J. T. High molecular weight precursor polypeptides to structural proteins of Rauscher murine leukemia virus. J Mol Biol. 1976 Nov 15;107(4):459–477. doi: 10.1016/s0022-2836(76)80078-2. [DOI] [PubMed] [Google Scholar]
  32. Strand M., August J. T. Structural proteins of oncogenic ribonucleic acid viruses. Interspec II, a new interspecies antigen. J Biol Chem. 1973 Aug 25;248(16):5627–5633. [PubMed] [Google Scholar]
  33. Thiel H. J., Beug H., Graf T., Schwarz H., Schäfer W., Bergholz C., Deinhardt F. Studies of simian sarcoma and simian sarcoma-associated virus. II. Isolation of the major viral glycoprotein, properties of this component and its specific antiserum. Virology. 1978 Oct 15;90(2):360–365. doi: 10.1016/0042-6822(78)90320-3. [DOI] [PubMed] [Google Scholar]
  34. Valentine R. C., Shapiro B. M., Stadtman E. R. Regulation of glutamine synthetase. XII. Electron microscopy of the enzyme from Escherichia coli. Biochemistry. 1968 Jun;7(6):2143–2152. doi: 10.1021/bi00846a017. [DOI] [PubMed] [Google Scholar]
  35. Van Zaane D., Dekker-Michielsen J. A., Bloemers H. P. Virus-specific precursor polypeptides in cells infected with Rauscher leukemia virus: synthesis, identification, and processing. Virology. 1976 Nov;75(1):113–129. doi: 10.1016/0042-6822(76)90011-8. [DOI] [PubMed] [Google Scholar]
  36. Witte O. N., Wirth D. F. Structure of the murine leukemia virus envelope glycoprotein precursor. J Virol. 1979 Feb;29(2):735–743. doi: 10.1128/jvi.29.2.735-743.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Witter R., Frank H., Moennig V., Hunsmann G., Lange J., Schäfer W. Properties of mouse leukemia viruses. IV. Hemagglutination assay and characterization of hemagglutinating surface components. Virology. 1973 Aug;54(2):330–345. doi: 10.1016/0042-6822(73)90147-5. [DOI] [PubMed] [Google Scholar]
  38. Zaane D. V., Gielkens A. L., Hesselink W. G., Bloemers H. P. Identification of Rauscher murine leukemia virus-specific mRNAs for the synthesis of gag- and env-gene products. Proc Natl Acad Sci U S A. 1977 May;74(5):1855–1859. doi: 10.1073/pnas.74.5.1855. [DOI] [PMC free article] [PubMed] [Google Scholar]

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