A novel type-1 ribosome-inactivating protein isolated from the supernatant of transformed suspension cultures of Trichosanthes kirilowii |
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Authors: | N.-J. Remi Shih K. A. McDonald A. M. Dandekar T. Girbés R. Iglesias A. P. Jackman |
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Affiliation: | (1) Department of Chemical Engineering and Materials Science, University of California, Davis, CA 95616, USA Fax no.: +1-916-752-1031 E-mail: kamcdonald@ucdavis.edu, HM;(2) Department of Pomology, University of California, Davis, CA 95616, USA, US;(3) Departmento de Bioquímica y Biología Molecular, Faultad de Ciencias, Universidad de Valladolid, E-47005 Valladolid, Spain, ES |
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Abstract: | A type-1 ribosome-inactivating protein (RIP) designated TK-35 has been purified from the supernatant of suspension cultures of Agrobacterium rhizogenes-transformed stem sections of Trichosanthes kirilowii. The protein was purified from the supernatant by PerSeptive SH/M cation exchange and Sephadex G-75 S gel permeation chromatography. The protein occurs as a monomer, with a molecular weight of 35,117, and is glycosylated. A protein translation inhibition assay indicates that TK-35 has an IC50 value of 2.45 nm and is able to release the rRNA N-glycosidase diagnostic fragment from rabbit reticulocyte lysate. TK-35 is quite thermally stable. Analysis of its N-terminal sequence and two lys-C-protease-digested polypeptides (internal) amino acid sequence indicates that this protein is not homologous to trichosanthin and other type-1 RIPs in Cucurbitaceae family. Received: 20 August 1997 / Revision received: 30 September 1997 / Accepted: 11 November 1997 |
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Keywords: | Ribosome-inactivating proteins Trichosanthes kirilowii Suspension culture |
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