Author: Kim Sung Yun Valencia Marcela Lee Eui Seung Park Daeho Oh Myungsok Xue Ding Park Woo Jin
Publisher: Humana Press, Inc
ISSN: 1073-6085
Source: Molecular Biotechnology, Vol.27, Iss.1, 2004-05, pp. : 1-6
Disclaimer: Any content in publications that violate the sovereignty, the constitution or regulations of the PRC is not accepted or approved by CNPIEC.
Abstract
Identifying cellular substrates repertoire of individual proteases will facilitate our understanding of their physiological and pathological roles. In this article, we employed a yeast-based screening method to isolate CED-3 substrates. This method uses a transcription factor anchored to the plasma membrane by fusion to a library of cellular protein sequences. When a fusion protein is cleaved by CED-3, the transcription factor is released from the plasma membrane and enters the nucleus where it turns on the expression of reporter genes. We identified seven candidate clones by screening a genomic library using this method. Of these seven clones, two were cleaved by purified CED-3 in vitro. Therefore, the method described here may be generally used for genomewide screening to isolate potential substrates of specific proteases.
Related content
A drug screening program for ion channels expressed in yeast
By Hahnenberger K.M. Kurtz S.E.
Trends in Biotechnology, Vol. 15, Iss. 1, 1997-01 ,pp. :