Binding method for FK-506- and rapamycin-like drugs with a novel

Chemistry: analytical and immunological testing – Biospecific ligand binding assay – Utilizing isolate of tissue or organ as binding agent

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436504, 436531, 436541, 436542, 436815, G01N 33538, G01N 33545

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active

055255230

ABSTRACT:
A new homogeneous cytosolic binding protein (FKBP-14.6 immunophilin) having a molecular weight of about 14.6 kDa (calculated from its amino acid composition), reversibly binds the immunosuppressive drugs FK-506, rapamycin or chemically related compounds, but not cyclosporine. The FKBP-14.6 protein has a pI of 6.5-7.5, and the (partial) N-terminal amino acid sequence: NH.sub.2 -Lys-Leu-Pro-Tyr-Glu-Leu-Lys-X-Asn-Val-Lys-Ala-Phe-X-X-Lys-Val- (where X is undefined), and partial internal amino acid sequences -Val-Leu-Asp-Thr-Ala-Tyr-Glu-Tyr-Gly-Ala-Glu-Ala-Leu-Glu-, -Glu-Phe-Thr-Pro-Val-Phe-Gln-Ala-X-Phe- and -Ser-Leu-Val-Pro-Leu-Val-Gly-X-Lys- (where X is undefined). This N-terminal amino acid sequence exhibits no homology to FKBP-12 or membrane-associated FKBP-13. THe FKBP-14.6 is isolated from the cytosol of mammalian tissues, preferably calf thymus, and can be used in diagnostic and purification procedures involving FK-506 and rapamycin-type immunosuppressant drugs.

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Jin et al., "Molecular Cloning of a 25-kDa High Affinity Rapamycin Binding Protein, FKBP25", J. Biol. Chem., 267:10942-945 (1992).
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