Genetic transfer of endothelin converting enzyme activity to CHO‐K1 cells: detection of positive cells by reverse hemolytic plaque assay

FEBS Letters

Ikura, Tsuyoshi¹  Sawamura, Tatsuya¹  Masaki, Tomoh¹  Miwa, Soichi¹  Shiraki, Takuma¹  Kobayashi, Shigeo² 

[1] Department of Pharmacology, Faculty of Medicine, Kyoto University, Kyoto 606, Japan;Department of Neuroscience, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto 606, Japan

关键词: Endothelin-1;    Endothelin converting enzyme;    Reverse hemolytic plaque assay;    ET-1;    endothelin-1;    big ET-1;    big endothelin-1;    ECE;    endotheline converting enzyme;   

DOI  :  10.1016/0014-5793(94)00830-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have established a novel method of molecular cloning of endothelin converting enzyme, a key enzyme in the production of a potent vasoconstrictor endothelin-1, by modification of the reverse hemolytic plaque assay. Also, we demonstrated that a cell line, CHO-K1, showed no detectable activity of endothelin converting enzyme. This cell line was transfected with a cDNA library of bovine endothelial cells. The modified reverse hemolytic plaque assay was shown to detect even a single CHO-K 1 cell that was changed to produce mature ET-1 by transfection. Thus, this novel method is suggested to be useful for the molecular cloning of other secreted antigens and their processing enzyme.

【 授权许可】

Unknown