Purification of polyoma virus medium-size tumor antigen by immunoaffinity chromatography.

Abstract

We have used antibodies against the synthetic peptide Lys-Arg-Ser-Arg-His-Phe, corresponding to the six COOH-terminal amino acids of the polyoma virus medium tumor (T) antigen, to purify the medium T antigen by affinity chromatography. Release of the medium T antigen from the anti-peptide antibody was achieved under mild conditions by using a large excess of the peptide in an isotonic buffer at neutral pH containing mixed detergents. This procedure yielded a 2,500-fold purification of the medium T antigen in a single step. The protein kinase activity associated with the medium T antigen was also released and was studied in this active state in solution. Sedimentation analysis showed that the bulk of the purified medium T antigen was in a monomeric form (Mr about 42,000) not associated with protein kinase activity. A small fraction of the medium T antigen was found in a rapidly sedimenting form (Mr about 200,000) that possessed protein kinase activity.

• Authors: G Walter, M A Hutchinson, T Hunter, W Eckhart
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 79 Issue 13 Pg. 4025-9 (Jul 1982) ISSN: 0027-8424 [Print] United States
• PMID: 6287461 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antibodies
• Antigen-Antibody Complex
• Antigens, Neoplasm
• Protein Kinases

Topics

• Animals
• Antibodies
• Antigen-Antibody Complex
• Antigens, Neoplasm (isolation & purification)
• Cells, Cultured
• Chromatography, Affinity
• Mice
• Molecular Weight
• Polyomavirus (enzymology, immunology)
• Protein Kinases (isolation & purification)