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Science 2 November 1984: Vol. 226. no. 4674, pp. 544 - 547 DOI: 10.1126/science.6238408
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Articles
Science, Vol 226, Issue 4674, 544-547
Copyright © 1984 by American Association for the Advancement of Science
Cyclophilin: a specific cytosolic binding protein for cyclosporin A
RE Handschumacher,
MW Harding,
J Rice,
RJ Drugge,
and
DW Speicher
Cyclophilin, a specific cytosolic binding protein responsible for the concentration of the immunosuppressant cyclosporin A by lymphoid cells, was purified to homogeneity from bovine thymocytes. Cation-exchange high-performance liquid chromatography resolved a major and minor cyclophilin species that bind cyclosporin A with a dissociation constant of about 2 X 10(-7) moles per liter and specific activities of 77 and 67 micrograms per milligram of protein, respectively. Both cyclophilin species have an apparent molecular weight of 15,000, an isoelectric point of 9.6, and nearly identical amino acid compositions. A portion of the NH2-terminal amino acid sequence of the major species was determined. The cyclosporin A-binding activity of cyclophilin is sulfhydryl dependent, unstable at 56 degrees C and at pH 4 or 9.5, and sensitive to trypsin but not to chymotrypsin digestion. Cyclophilin specifically binds a series of cyclosporin analogs in proportion to their activity in a mixed lymphocyte reaction. Isolation of cyclophilin from the cytosol of thymocytes suggests that the immunosuppressive activity of cyclosporin A is mediated by an intracellular mechanism, not by a membrane-associated mechanism.
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Science
253, 905-909
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Science
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Science
250, 556-559
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Science
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