Partial purification of the Epstin-Barr virus nuclear antigen(s)

T. B. Sculley, T. Kreofsky, G. R. Pearson, T. C. Spelsberg

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The Epstein-Barr virus nuclear antigen (EBNA) is speculated to be involved in cell transformation by the virus. Studies on the molecular properties of EBNA, however, have yielded conflicting results. In this study, three Epstein-Barr virus- (EBV)-induced antigens were isolated and purified from extracts prepared from Raji cells. These antigens were able to block the anticomplement immunofluorescence reaction, indicating that all three were related to EBNA. The soluble antigen was found wholly in the cytosol fraction. An EBV-induced nuclear antigen I was found both in the cytosol and the nucleus. The EBV-induced nuclear antigen II was found associated with the chromatin. The soluble antigen and the nuclear antigen I were separated and partially purified using phosphocellulose chromatography. Each was further purified 1,400-fold with respect to the whole cell state by chromatography on CL-Sepharose 6B followed by blue dextran-Sepharose. Subunit molecular weights of 70,000 were determined for each of these antigens, both in the crude and purified state, by radioimmunoelectrophoresis and gel filtration. The nuclear antigen II was purified 2,500-fold using hydroxylapatite, CL-Sepharose 6B, and blue dextran-Sepharose chromatographies. This antigen displayed two subunits by radioimmunoelectrophoresis with molecular weights of 65,000 and 70,000. Although all antigens shared similar molecular weights, the extent of their homology remains to be determined.

Original languageEnglish (US)
Pages (from-to)3974-3982
Number of pages9
JournalJournal of Biological Chemistry
Volume258
Issue number6
StatePublished - 1983

Fingerprint

Nuclear Antigens
Epstein-Barr Virus Nuclear Antigens
Viruses
Purification
Antigens
Chromatography
Molecular weight
Molecular Weight
Cytosol
Agarose Chromatography
Durapatite
Chromatin
Human Herpesvirus 4
Gel Chromatography
Fluorescent Antibody Technique
Gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sculley, T. B., Kreofsky, T., Pearson, G. R., & Spelsberg, T. C. (1983). Partial purification of the Epstin-Barr virus nuclear antigen(s). Journal of Biological Chemistry, 258(6), 3974-3982.

Partial purification of the Epstin-Barr virus nuclear antigen(s). / Sculley, T. B.; Kreofsky, T.; Pearson, G. R.; Spelsberg, T. C.

In: Journal of Biological Chemistry, Vol. 258, No. 6, 1983, p. 3974-3982.

Research output: Contribution to journalArticle

Sculley, TB, Kreofsky, T, Pearson, GR & Spelsberg, TC 1983, 'Partial purification of the Epstin-Barr virus nuclear antigen(s)', Journal of Biological Chemistry, vol. 258, no. 6, pp. 3974-3982.
Sculley TB, Kreofsky T, Pearson GR, Spelsberg TC. Partial purification of the Epstin-Barr virus nuclear antigen(s). Journal of Biological Chemistry. 1983;258(6):3974-3982.
Sculley, T. B. ; Kreofsky, T. ; Pearson, G. R. ; Spelsberg, T. C. / Partial purification of the Epstin-Barr virus nuclear antigen(s). In: Journal of Biological Chemistry. 1983 ; Vol. 258, No. 6. pp. 3974-3982.
@article{041e17b2459846f9bb6bbc7824f0c6b0,
title = "Partial purification of the Epstin-Barr virus nuclear antigen(s)",
abstract = "The Epstein-Barr virus nuclear antigen (EBNA) is speculated to be involved in cell transformation by the virus. Studies on the molecular properties of EBNA, however, have yielded conflicting results. In this study, three Epstein-Barr virus- (EBV)-induced antigens were isolated and purified from extracts prepared from Raji cells. These antigens were able to block the anticomplement immunofluorescence reaction, indicating that all three were related to EBNA. The soluble antigen was found wholly in the cytosol fraction. An EBV-induced nuclear antigen I was found both in the cytosol and the nucleus. The EBV-induced nuclear antigen II was found associated with the chromatin. The soluble antigen and the nuclear antigen I were separated and partially purified using phosphocellulose chromatography. Each was further purified 1,400-fold with respect to the whole cell state by chromatography on CL-Sepharose 6B followed by blue dextran-Sepharose. Subunit molecular weights of 70,000 were determined for each of these antigens, both in the crude and purified state, by radioimmunoelectrophoresis and gel filtration. The nuclear antigen II was purified 2,500-fold using hydroxylapatite, CL-Sepharose 6B, and blue dextran-Sepharose chromatographies. This antigen displayed two subunits by radioimmunoelectrophoresis with molecular weights of 65,000 and 70,000. Although all antigens shared similar molecular weights, the extent of their homology remains to be determined.",
author = "Sculley, {T. B.} and T. Kreofsky and Pearson, {G. R.} and Spelsberg, {T. C.}",
year = "1983",
language = "English (US)",
volume = "258",
pages = "3974--3982",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "6",

}

TY - JOUR

T1 - Partial purification of the Epstin-Barr virus nuclear antigen(s)

AU - Sculley, T. B.

AU - Kreofsky, T.

AU - Pearson, G. R.

AU - Spelsberg, T. C.

PY - 1983

Y1 - 1983

N2 - The Epstein-Barr virus nuclear antigen (EBNA) is speculated to be involved in cell transformation by the virus. Studies on the molecular properties of EBNA, however, have yielded conflicting results. In this study, three Epstein-Barr virus- (EBV)-induced antigens were isolated and purified from extracts prepared from Raji cells. These antigens were able to block the anticomplement immunofluorescence reaction, indicating that all three were related to EBNA. The soluble antigen was found wholly in the cytosol fraction. An EBV-induced nuclear antigen I was found both in the cytosol and the nucleus. The EBV-induced nuclear antigen II was found associated with the chromatin. The soluble antigen and the nuclear antigen I were separated and partially purified using phosphocellulose chromatography. Each was further purified 1,400-fold with respect to the whole cell state by chromatography on CL-Sepharose 6B followed by blue dextran-Sepharose. Subunit molecular weights of 70,000 were determined for each of these antigens, both in the crude and purified state, by radioimmunoelectrophoresis and gel filtration. The nuclear antigen II was purified 2,500-fold using hydroxylapatite, CL-Sepharose 6B, and blue dextran-Sepharose chromatographies. This antigen displayed two subunits by radioimmunoelectrophoresis with molecular weights of 65,000 and 70,000. Although all antigens shared similar molecular weights, the extent of their homology remains to be determined.

AB - The Epstein-Barr virus nuclear antigen (EBNA) is speculated to be involved in cell transformation by the virus. Studies on the molecular properties of EBNA, however, have yielded conflicting results. In this study, three Epstein-Barr virus- (EBV)-induced antigens were isolated and purified from extracts prepared from Raji cells. These antigens were able to block the anticomplement immunofluorescence reaction, indicating that all three were related to EBNA. The soluble antigen was found wholly in the cytosol fraction. An EBV-induced nuclear antigen I was found both in the cytosol and the nucleus. The EBV-induced nuclear antigen II was found associated with the chromatin. The soluble antigen and the nuclear antigen I were separated and partially purified using phosphocellulose chromatography. Each was further purified 1,400-fold with respect to the whole cell state by chromatography on CL-Sepharose 6B followed by blue dextran-Sepharose. Subunit molecular weights of 70,000 were determined for each of these antigens, both in the crude and purified state, by radioimmunoelectrophoresis and gel filtration. The nuclear antigen II was purified 2,500-fold using hydroxylapatite, CL-Sepharose 6B, and blue dextran-Sepharose chromatographies. This antigen displayed two subunits by radioimmunoelectrophoresis with molecular weights of 65,000 and 70,000. Although all antigens shared similar molecular weights, the extent of their homology remains to be determined.

UR - http://www.scopus.com/inward/record.url?scp=0020643797&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020643797&partnerID=8YFLogxK

M3 - Article

VL - 258

SP - 3974

EP - 3982

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 6

ER -