Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1976 Mar;17(3):906–915. doi: 10.1128/jvi.17.3.906-915.1976

Analysis of structural proteins of purified murine cytomegalovirus.

K S Kim, V J Sapienza, R I Carp, H M Moon
PMCID: PMC515491  PMID: 176450

Abstract

Murine cytomegalovirus propagated in mouse embryo fibroblasts was purified by the following procedures. (i) Extracellular virus was concentrated by centrifugation at 100,000 x g for 90 min. (ii) The concentrated virus was passed through a Bio-Rad Bio-Gel A-15m column to eliminate contaminating materials smaller than 15 x 10(6) daltons. Most of the virus was recovered in the void volume of the column. (iii) Two consecutive centrifugations through 20 to 50% potassium tartrate gradients were performed. After the second tartrate gradient centrifugation, symmetrical, coinciding peaks of plaque titer, protein, and radioactivity were found at a density between 1.20 g/cm3 and 1.21 g/cm3. To establish purification criteria, virus was purified from two different mixtures: [35S]methionine-labeled extracellular virus, mixed with an equal volume of unlabeled normal culture fluid, and unlabeled extracellular virus mixed with an equal volume of [35S]methionine-labeled normal culture fluid. At the end of the procedure, the extent of purification, as judged by the ratio of cellular to viral radioactivity was at least 70-fold. Virus proteins were analyzed by electrophoresis on a 5 to 20% gradient polyacrylamide gel slab. After gel electrophoresis,, Coomassie brilliant blue staining profiles and autoradiograms of the purified virus preparations were compared. At least 33 virus structural protein bands were present. The molecular weights of these proteins ranged from 11,500 to 255,000. The sum of the molecular weights of the virus structural proteins was 2,462,000. Autoradiograms obtained from electrophoresis of purified [14C]glucosamine-labeled virus showed that at lease 6 of the 33 viral structural proteins were glycoproteins.

Full text

PDF
915

Images in this article

911Image
on p.911
914Image
on p.914

Selected References

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

  1. Abodeely R. A., Lawson L. A., Randall C. C. Morphology and entry of enveloped and deenveloped equine abortion (herpes) virus. J Virol. 1970 Apr;5(4):513–523. doi: 10.1128/jvi.5.4.513-523.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abodeely R. A., Palmer E., Lawson L. A., Randall C. C. The proteins of enveloped and deenveloped equine abortion (herpes) virus and the separated envelope. Virology. 1971 Apr;44(1):146–152. doi: 10.1016/0042-6822(71)90161-9. [DOI] [PubMed] [Google Scholar]
  3. Ben-Porat T., Shimono H., Kaplan A. S. Synthesis of proteins in cells infected with herpesvirus. IV. Analysis of the proteins in viral particles isolated from the cytoplasm and the nucleus. Virology. 1970 Jun;41(2):256–264. doi: 10.1016/0042-6822(70)90077-2. [DOI] [PubMed] [Google Scholar]
  4. Chen J. H., Lee L. F., Nazerian K., Burmester B. R. Structural proteins of Marek's disease virus. Virology. 1972 Feb;47(2):434–443. doi: 10.1016/0042-6822(72)90279-6. [DOI] [PubMed] [Google Scholar]
  5. Heine J. W., Honess R. W., Cassai E., Roizman B. Proteins specified by herpes simplex virus. XII. The virion polypeptides of type 1 strains. J Virol. 1974 Sep;14(3):640–651. doi: 10.1128/jvi.14.3.640-651.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaplan A. S., Ben-Porat T. Synthesis of proteins in cells infected with herpesvirus, VI. Characterization of the proteins of the viral membrane. Proc Natl Acad Sci U S A. 1970 Jul;66(3):799–806. doi: 10.1073/pnas.66.3.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kaplan A. S., Shimono H., Ben-Porat T. Synthesis of proteins in cells infected with herpesvirus. 3. Relative amino acid content of various proteins formed after infection. Virology. 1970 Jan;40(1):90–101. doi: 10.1016/0042-6822(70)90382-x. [DOI] [PubMed] [Google Scholar]
  8. Kemp M. C., Perdue M. L., Rogers H. W., O'Callaghan D. J., Randall C. C. Structural polypeptides of the hamster strain of equine herpes virus type 1: products associated with purification. Virology. 1974 Oct;61(2):361–375. doi: 10.1016/0042-6822(74)90274-8. [DOI] [PubMed] [Google Scholar]
  9. Kim K. S., Carp R. I. Abortive infection of human diploid cells by murine cytomegalovirus. Infect Immun. 1972 Nov;6(5):793–797. doi: 10.1128/iai.6.5.793-797.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kim K. S., Carp R. I. Growth of murine cytomegalovirus in various cell lines. J Virol. 1971 Jun;7(6):720–725. doi: 10.1128/jvi.7.6.720-725.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Mosmann T. R., Hudson J. B. Some properties of the genome of murine cytomegalovirus (MCV). Virology. 1973 Jul;54(1):135–149. doi: 10.1016/0042-6822(73)90123-2. [DOI] [PubMed] [Google Scholar]
  13. Nazerian K., Lindahl T., Klein G., Lee L. F. Deoxyribonucleic acid of Marek's disease virus in virus-induced tumors. J Virol. 1973 Oct;12(4):841–846. doi: 10.1128/jvi.12.4.841-846.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  15. Shimono H., Ben-Porat T., Kaplan A. S. Synthesis of proteins in cells infected with herpesvirus. I. Structural viral proteins. Virology. 1969 Jan;37(1):49–55. doi: 10.1016/0042-6822(69)90305-5. [DOI] [PubMed] [Google Scholar]
  16. Spear P. G., Roizman B. Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpesvirion. J Virol. 1972 Jan;9(1):143–159. doi: 10.1128/jvi.9.1.143-159.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Sydiskis R. J., Roizman B. Polysomes and protein synthesis in cells infected with a DNA virus. Science. 1966 Jul 1;153(3731):76–78. doi: 10.1126/science.153.3731.76. [DOI] [PubMed] [Google Scholar]
  19. 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]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES