Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1979 Sep;31(3):696–706. doi: 10.1128/jvi.31.3.696-706.1979

Characterization of an immune complex kinase in immunoprecipitates of avian sarcoma virus-transformed fibroblasts.

N D Richert, P J Davies, G Jay, I H Pastan
PMCID: PMC353498  PMID: 41953

Abstract

Kinase activity detected in immune complexes containing the src gene product of the avian sarcoma virus has been reported. To further characterize this immune complex kinase, we developed a routine quantitative assay involving trichloroacetic acid precipitation on filters. The enzyme reaction required either Mg2+ or Mn2+, but was inactive with Ca2+. The kinetics of the phosphorylation reaction indicated a transient enzyme activity limited by rapid substrate-dependent inactivation of the enzyme. A variety of nucleoside and deoxyribonucleoside triphosphates (dATP, ATP, GTP, CTP, dGTP, TTP, dCTP) served as phosphoryl donors. The phosphorylation of immunoglobulin G was inhibited by the presence of nucleoside diphosphates. Deoxyribonucleoside diphosphates can either stimulate or inhibit the kinase reaction depending upon the concentration used. The unusual enzymatic properties of the immune complex kinase raise the possibility that the enzyme does not function as a protein kinase in vivo, but rather belongs to a different class of kinases (nucleotide kinases) which adventitiously phosphorylates immunoglobulin G when immunoprecipitated with immune serum.

Full text

PDF
695

Images in this article

Selected References

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

  1. Agren G., Pontén J., Ronquist G., Westermark B. Nucleoside diphosphate kinase at the cell surface of neoplastic human cells in culture. J Cell Physiol. 1974 Feb;83(1):91–101. doi: 10.1002/jcp.1040830113. [DOI] [PubMed] [Google Scholar]
  2. Bingham E. W., Farrel H. M., Jr Casein kinase from the Golgi apparatus of lactating mammary gland. J Biol Chem. 1974 Jun 10;249(11):3647–3651. [PubMed] [Google Scholar]
  3. Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
  4. Collett M. S., Brugge J. S., Erikson R. L. Characterization of a normal avian cell protein related to the avian sarcoma virus transforming gene product. Cell. 1978 Dec;15(4):1363–1369. doi: 10.1016/0092-8674(78)90061-2. [DOI] [PubMed] [Google Scholar]
  5. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goldstein J. L., Hasty M. A. Phosvitin kinase from the liver of the rooster. Purification and partial characterization. J Biol Chem. 1973 Sep 25;248(18):6300–6307. [PubMed] [Google Scholar]
  7. Hatanaka M., Twiddy E., Gilden R. V. Protein kinase associated with RNA tumor viruses and other budding RNA viruses. Virology. 1972 Feb;47(2):536–538. doi: 10.1016/0042-6822(72)90297-8. [DOI] [PubMed] [Google Scholar]
  8. Ives D. H., Durham J. P. Deoxycytidine kinase. 3. Kinetics and allosteric regulation of the calf thymus enzyme. J Biol Chem. 1970 May 10;245(9):2285–2294. [PubMed] [Google Scholar]
  9. Jay G., Shiu R. P., Jay F. T., Levine A. S., Pastan I. Identification of a transformation-specific protein induced by a Rous sarcoma virus. Cell. 1978 Mar;13(3):527–534. doi: 10.1016/0092-8674(78)90326-4. [DOI] [PubMed] [Google Scholar]
  10. Junghans R. P., Hu S., Knight C. A., Davidson N. Heteroduplex analysis of avian RNA tumor viruses. Proc Natl Acad Sci U S A. 1977 Feb;74(2):477–481. doi: 10.1073/pnas.74.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Kishimoto A., Takai Y., Nishizuka Y. Activation of glycogen phosphorylase kinase by a calcium-activated, cyclic nucleotide-independent protein kinase system. J Biol Chem. 1977 Nov 10;252(21):7449–7452. [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Lassam N. J., Bayley S. T., Graham F. L., Branton P. E. Immunoprecipitation of protein kinase activity from adenovirus 5-infected cells using antiserum directed against tumour antigens. Nature. 1979 Jan 18;277(5693):241–243. doi: 10.1038/277241a0. [DOI] [PubMed] [Google Scholar]
  16. Levinson A. D., Oppermann H., Levintow L., Varmus H. E., Bishop J. M. Evidence that the transforming gene of avian sarcoma virus encodes a protein kinase associated with a phosphoprotein. Cell. 1978 Oct;15(2):561–572. doi: 10.1016/0092-8674(78)90024-7. [DOI] [PubMed] [Google Scholar]
  17. OKAZAKI R., KORNBERG A. DEOXYTHYMIDINE KINASE OF ESCHERICHIA COLI. II. KINETICS AND FEEDBACK CONTROL. J Biol Chem. 1964 Jan;239:275–284. [PubMed] [Google Scholar]
  18. Pires E. M., Perry S. V. Purification and properties of myosin light-chain kinase from fast skeletal muscle. Biochem J. 1977 Oct 1;167(1):137–146. doi: 10.1042/bj1670137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Purchio A. F., Erikson E., Brugge J. S., Erikson R. L. Identification of a polypeptide encoded by the avian sarcoma virus src gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1567–1571. doi: 10.1073/pnas.75.3.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Purchio A. F., Erikson E., Erikson R. L. Translation of 35S and of subgenomic regions of avian sarcoma virus RNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4661–4665. doi: 10.1073/pnas.74.10.4661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rosok M. J., Watson K. F. Fractionation of two protein kinases from avian myeloblastosis virus and characterization of the protein kinase activity preferring basic phosphoacceptor proteins. J Virol. 1979 Mar;29(3):872–880. doi: 10.1128/jvi.29.3.872-880.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sefton B. M., Beemon K., Hunter T. Comparison of the expression of the src gene of Rous sarcoma virus in vitro and in vivo. J Virol. 1978 Dec;28(3):957–971. doi: 10.1128/jvi.28.3.957-971.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shiu R. P., Pouyssegur J., Pastan I. Glucose depletion accounts for the induction of two transformation-sensitive membrane proteinsin Rous sarcoma virus-transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3840–3844. doi: 10.1073/pnas.74.9.3840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Silberstein H., August J. T. Purification and properties of a virion protein kinase. J Biol Chem. 1976 May 25;251(10):3176–3184. [PubMed] [Google Scholar]
  25. Spector D. H., Smith K., Padgett T., McCombe P., Roulland-Dussoix D., Moscovici C., Varmus H. E., Bishop J. M. Uninfected avian cells contain RNA related to the transforming gene of avian sarcoma viruses. Cell. 1978 Feb;13(2):371–379. doi: 10.1016/0092-8674(78)90205-2. [DOI] [PubMed] [Google Scholar]
  26. Stehelin D., Varmus H. E., Bishop J. M., Vogt P. K. DNA related to the transforming gene(s) of avian sarcoma viruses is present in normal avian DNA. Nature. 1976 Mar 11;260(5547):170–173. doi: 10.1038/260170a0. [DOI] [PubMed] [Google Scholar]
  27. Takai Y., Kishimoto A., Inoue M., Nishizuka Y. Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues. I. Purification and characterization of an active enzyme from bovine cerebellum. J Biol Chem. 1977 Nov 10;252(21):7603–7609. [PubMed] [Google Scholar]
  28. Traugh J. A., Traut R. R. Characterization of protein kinases from rabbit reticulocytes. J Biol Chem. 1974 Feb 25;249(4):1207–1212. [PubMed] [Google Scholar]
  29. Weiss S. R., Varmus H. E., Bishop J. M. The size and genetic composition of virus-specific RNAs in the cytoplasm of cells producing avian sarcoma-leukosis viruses. Cell. 1977 Dec;12(4):983–992. doi: 10.1016/0092-8674(77)90163-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES