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. 1978 Aug;27(2):436–442. doi: 10.1128/jvi.27.2.436-442.1978

Chromatographic separation of the polyoma virus proteins and renaturation of the isolated VP1 major capsid protein.

J N Brady, R A Consigli
PMCID: PMC354182  PMID: 211269

Abstract

Treatment of purified polyoma virions with 6 M guanidine-hydrochloride and 0.01 M beta-mercaptoethanol resulted in the immediate loss of both hemagglutinating and plaque-forming ability. Gel filtration through Sepharose CL-6B beads allowed separation of the dimer, VP1, VP2, VP3, and histone proteins VP4-7 in highly purified form. Renaturation of the purified VP1 protein resulted in the formation of subunits that were morphologically, biophysically, and immunologically similar to native virion capsomeres.

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Selected References

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

  1. Brady J. N., Winston V. D., Consigli R. A. Characterization of a DNA-protein complex and capsomere subunits derived from polyoma virus by treatment with ethyleneglycol-bis-N,N'-tetraacetic acid and dithiothreitol. J Virol. 1978 Jul;27(1):193–204. doi: 10.1128/jvi.27.1.193-204.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brady J. N., Winston V. D., Consigli R. A. Dissociation of polyoma virus by the chelation of calcium ions found associated with purified virions. J Virol. 1977 Sep;23(3):717–724. doi: 10.1128/jvi.23.3.717-724.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bryce C. F., Crichton R. R. The subunit structure of horse spleen apoferritin. I. The molecular weight of the subunit. J Biol Chem. 1971 Jul 10;246(13):4198–4205. [PubMed] [Google Scholar]
  4. Consigli R. A., Zabielski J., Weil R. Plaque assay for polyoma virus on primary mouse kidney cell cultures. Appl Microbiol. 1973 Oct;26(4):627–628. doi: 10.1128/am.26.4.627-628.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fey G., Hirt B. Fingerprints of polyoma virus proteins and mouse histones. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):235–241. doi: 10.1101/sqb.1974.039.01.030. [DOI] [PubMed] [Google Scholar]
  6. Fish W. W., Mann K. G., Tanford C. The estimation of polypeptide chain molecular weights by gel filtration in 6 M guanidine hydrochloride. J Biol Chem. 1969 Sep 25;244(18):4989–4994. [PubMed] [Google Scholar]
  7. Frearson P. M., Crawford L. V. Polyoma virus basic proteins. J Gen Virol. 1972 Feb;14(2):141–155. doi: 10.1099/0022-1317-14-2-141. [DOI] [PubMed] [Google Scholar]
  8. Frost E., Bourgaux P. Decapsidation of polyoma virus: identification of subviral species. Virology. 1975 Nov;68(1):245–255. doi: 10.1016/0042-6822(75)90165-8. [DOI] [PubMed] [Google Scholar]
  9. Gibson W. Polyoma virus proteins: a description of the structural proteins of the virion based on polyacrylamide gel electrophoresis and peptide analysis. Virology. 1974 Dec;62(2):319–336. doi: 10.1016/0042-6822(74)90395-x. [DOI] [PubMed] [Google Scholar]
  10. Hewick R. M., Fried M., Waterfield M. D. Nonhistone virion proteins of polyoma: characterisation of the particle proteins by tryptic peptide analysis by use of ion-exchange columns. Virology. 1975 Aug;66(2):408–419. doi: 10.1016/0042-6822(75)90213-5. [DOI] [PubMed] [Google Scholar]
  11. Kekwick R. A. The serum proteins in multiple myelomatosis. Biochem J. 1940 Sep;34(8-9):1248–1257. doi: 10.1042/bj0341248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LEVY H. B., SOBER H. A. A simple chromatographic method for preparation of gamma globulin. Proc Soc Exp Biol Med. 1960 Jan;103:250–252. doi: 10.3181/00379727-103-25476. [DOI] [PubMed] [Google Scholar]
  13. Mackay R. L., Consigli R. A. Early events in polyoma virus infection: attachment, penetration, and nuclear entry. J Virol. 1976 Aug;19(2):620–636. doi: 10.1128/jvi.19.2.620-636.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McMillen J., Center M. S., Consigli R. A. Origin of the polyoma virus-associated endonuclease. J Virol. 1975 Jan;17(1):127–131. doi: 10.1128/jvi.17.1.127-131.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McMillen J., Consigli R. A. Characterization of polyoma DNA-protein complexes. I. Electrophoretic identification of the proteins in a nucleoprotein complex isolated from polyoma-infected cells. J Virol. 1974 Dec;14(6):1326–1336. doi: 10.1128/jvi.14.6.1326-1336.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McMillen J., Consigli R. A. Immunological reactivity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions. J Virol. 1977 Mar;21(3):1113–1120. doi: 10.1128/jvi.21.3.1113-1120.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McMillen J., Consigli R. A. In vitro radioisotopic labeling of proteins associated with purified polyoma virions. J Virol. 1974 Dec;14(6):1627–1629. doi: 10.1128/jvi.14.6.1627-1629.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roblin R., Härle E., Dulbecco R. Polyoma virus proteins. 1. Multiple virion components. Virology. 1971 Sep;45(3):555–566. doi: 10.1016/0042-6822(71)90171-1. [DOI] [PubMed] [Google Scholar]
  19. Smith G. L., Consigli R. A. Transient inhibition of polyoma virus synthesis by Sendai virus (parainfluenza I). I. Demonstration and nature of the inhibition by inactivated virus. J Virol. 1972 Dec;10(6):1091–1097. doi: 10.1128/jvi.10.6.1091-1097.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Walter G., Deppert W. Intermolecular disulfide bonds: an important structural feature of the polyoma virus capsid. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):255–257. doi: 10.1101/sqb.1974.039.01.033. [DOI] [PubMed] [Google Scholar]

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