从栝楼转化悬浮培养物的上清液中分离出的一种新型1型核糖体失活蛋白。

A novel type-1 ribosome-inactivating protein isolated from the supernatant of transformed suspension cultures of Trichosanthes kirilowii.

作者信息

Remi Shih N-J, McDonald K A, Dandekar A M, Girbés T, Iglesias R, Jackman A P

机构信息

Department of Chemical Engineering and Materials Science, University of California, Davis, CA 95616, USA Fax no.: +1-916-752-1031 E-mail:

Department of Pomology, University of California, Davis, CA 95616, USA, , , , , , US.

出版信息

Plant Cell Rep. 1998 Apr;17(6-7):531-537. doi: 10.1007/s002990050437.

DOI:10.1007/s002990050437

PMID:30736631

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 IC 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.

摘要

一种名为TK-35的1型核糖体失活蛋白（RIP）已从发根农杆菌转化的栝楼茎段悬浮培养物的上清液中纯化出来。该蛋白通过PerSeptive SH/M阳离子交换和Sephadex G-75 S凝胶渗透色谱从上清液中纯化得到。该蛋白以单体形式存在，分子量为35,117，且为糖基化蛋白。蛋白质翻译抑制试验表明，TK-35的IC值为2.45 nM，并且能够从兔网织红细胞裂解物中释放出rRNA N-糖苷酶诊断片段。TK-35具有相当高的热稳定性。对其N端序列和两个经赖氨酰内肽酶消化的多肽（内部）氨基酸序列的分析表明，该蛋白与天花粉蛋白以及葫芦科家族中的其他1型RIPs没有同源性。