Specific interaction of the cyclophilin-cyclosporin complex with the B subunit of calcineurin

Wei Li, R. E. Handschumacher

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Calcineurin (CaN), a Ca2+/calmodulin-dependent serine/threonine phosphatase, has been shown to be inhibited by the complex of the immunosuppressant cyclosporin A (CsA) and its receptor, cyclophilin (CyP), but not by either alone. In the current study, the topological relationship between cyclophilin and the subunits of CaN has been explored using chemical cross-linking agents. In the presence of cyclosporin, 125I-CyP, shown to bind to CsA, is extensively cross-linked to the B subunit of CaN but not the catalytic A subunit. However, the A subunit is required for binding of the CyP·CsA complex, since cross-linking to recombinant B subunit alone does not occur. The kinetics of association indicate a saturable reaction with a K(d) of less than 70 nM. Cross-linking to the B subunit occurs with cross-linkers that span from 0 to 16 Å and employ different cross-linking chemistry, indicating direct contact between the B subunit and CyP. Similar cross- linking to the B subunit has been observed with the complex of 125I- labeled FK506 binding protein (FKBP) and FK506 but not with the FKBP- rapamycin complex. CyP·CsA cross-linking to CaN is Ca2+/calmodulin- dependent with intact CaN, but Ca2+/calmodulin-independent after digestion to remove the calmodulin binding and autoinhibitory domain.

Original languageEnglish (US)
Pages (from-to)14040-14044
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number19
StatePublished - 1993
Externally publishedYes

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Cyclophilins
Calcineurin
Cyclosporine
Calmodulin
Tacrolimus Binding Proteins
Phosphoprotein Phosphatases
Tacrolimus
Sirolimus
Immunosuppressive Agents
Crosslinking
Digestion
Catalytic Domain
Association reactions
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Specific interaction of the cyclophilin-cyclosporin complex with the B subunit of calcineurin. / Li, Wei; Handschumacher, R. E.

In: Journal of Biological Chemistry, Vol. 268, No. 19, 1993, p. 14040-14044.

Research output: Contribution to journalArticle

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