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. 1976 Feb;73(2):520–523. doi: 10.1073/pnas.73.2.520

Interferon: purification and initial characterization from human diploid cells.

E Knight Jr
PMCID: PMC335941  PMID: 1061154

Abstract

Interferon produced by human diploid fibroblast cells in culture has been purified approximately 5000-fold. The purified interferon, when analyzed by electrophoresis on polyacrylamide gel containing sodium dodecyl sulfate, contains only one polypeptide component of 20,000 molecular weight. The interferon activity comigrates with this polypeptide, indicating identity of the activity and the polypeptide. Oxidation of this polypeptide with periodic acid and subsequent staining with Fuchsin base indicates that it contains carbohydrate ans suggests that the human fibroblast interferon is a glycoprotein.

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Selected References

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  1. Anfinsen C. B., Bose S., Corley L., Gurari-Rotman D. Partial purification of human interferon by affinity chromatography. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3139–3142. doi: 10.1073/pnas.71.8.3139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BUONASSISI V., SATO G., COHEN A. I. Hormone-producing cultures of adrenal and pituitary tumor origin. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1184–1190. doi: 10.1073/pnas.48.7.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Basch R. S., Goldstein G. Induction of T-cell differentiation in vitro by thymin, a purified polypeptide hormone of the thymus. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1474–1478. doi: 10.1073/pnas.71.4.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berg K., Ogburn C. A., Paucker K., Mogensen K. E., Cantell K. Affinity chromatography of human leukocyte and diploid cell interferons on sepharose-bound antibodies. J Immunol. 1975 Feb;114(2 Pt 1):640–644. [PubMed] [Google Scholar]
  5. CORNBLATH M., RANDLE P. J., PARMEGGIANI A., MORGAN H. E. Regulation of glycogenolysis in muscle. Effects of glucagon and anoxia on lactate production, glycogen content, and phosphorylase activity in the perfused isolated rat heart. J Biol Chem. 1963 May;238:1592–1597. [PubMed] [Google Scholar]
  6. Cohen S. The stimulation of epidermal proliferation by a specific protein (EGF). Dev Biol. 1965 Dec;12(3):394–407. doi: 10.1016/0012-1606(65)90005-9. [DOI] [PubMed] [Google Scholar]
  7. Frazier W. A., Boyd L. F., Bradshaw R. A. Interaction of nerve growth factor with surface membranes: biological competence of insolubilized nerve growth factor. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2931–2935. doi: 10.1073/pnas.70.10.2931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Havell E. A., Vilcek J., Falcoff E., Berman B. Suppression of human interferon production by inhibitors of glycosylation. Virology. 1975 Feb;63(2):475–483. doi: 10.1016/0042-6822(75)90320-7. [DOI] [PubMed] [Google Scholar]
  9. Havell E. A., Vilcek J. Production of high-titered interferon in cultures of human diploid cells. Antimicrob Agents Chemother. 1972 Dec;2(6):476–484. doi: 10.1128/aac.2.6.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huang J. W., Davey M. W., Hejna C. J., Von Muenchhausen W., Sulkowski E., Carter W. A. Selective binding of human interferon to albumin immoblized on agarose. J Biol Chem. 1974 Jul 25;249(14):4665–4667. [PubMed] [Google Scholar]
  11. Huang J. W., Davey M. W., Hejna C. J., Von Muenchhausen W., Sulkowski E., Carter W. A. Selective binding of human interferon to albumin immoblized on agarose. J Biol Chem. 1974 Jul 25;249(14):4665–4667. [PubMed] [Google Scholar]
  12. Kapitany R. A., Zebrowski E. J. A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal Biochem. 1973 Dec;56(2):361–369. doi: 10.1016/0003-2697(73)90202-9. [DOI] [PubMed] [Google Scholar]
  13. Knight E., Jr Heterogeneity of purified mouse interferons. J Biol Chem. 1975 Jun 10;250(11):4139–4144. [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Mayer M. M., Miller J. A. Photometric analysis of proteins and peptides at 191-194 millimicrons. Anal Biochem. 1970 Jul;36(1):91–100. doi: 10.1016/0003-2697(70)90335-0. [DOI] [PubMed] [Google Scholar]
  17. McGuire J. C., Pène J. J., Barrow-Carraway J. Gene expression during the development of bacteriophage phi 29. 3. Analysis of viral-specific protein synthesis with suppressible mutants. J Virol. 1974 Mar;13(3):690–698. doi: 10.1128/jvi.13.3.690-698.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pastan I., Roth J., Macchia V. Binding of hormone to tissue: the first step in polypeptide hormone action. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1802–1809. doi: 10.1073/pnas.56.6.1802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. RODBELL M. METABOLISM OF ISOLATED FAT CELLS. I. EFFECTS OF HORMONES ON GLUCOSE METABOLISM AND LIPOLYSIS. J Biol Chem. 1964 Feb;239:375–380. [PubMed] [Google Scholar]
  20. Stancek D., Gressnerová M., Paucker K. Isoelectric components of mouse, human, and rabbit interferons. Virology. 1970 Aug;41(4):740–750. doi: 10.1016/0042-6822(70)90438-1. [DOI] [PubMed] [Google Scholar]
  21. Stewart W. E., 2nd Distinct molecular species of interferons. Virology. 1974 Sep;61(1):80–86. doi: 10.1016/0042-6822(74)90243-8. [DOI] [PubMed] [Google Scholar]
  22. 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]

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