Rapid modification of proteins using a rapamycin-inducible tobacco etch virus protease system.

ABSTRACT:

Background

The ability to disrupt the function of a specific protein on a rapid time scale provides a powerful tool for biomedical research. Specific proteases provide a potential method to selectively cleave a chosen protein, but rapid control of protease activity is difficult.

Methodology/principal findings

A heterologous expression system for rapid target-directed proteolysis in mammalian cells was developed. The system consists of an inducible NIa protease from the tobacco etch virus (TEVp) and a chosen protein into which a TEVp substrate recognition sequence (TRS) has been inserted. Inducible activity was conferred to the TEVp using rapamycin-controlled TEVp fragment complementation. TEVp activity was assayed using a FRET-based reporter construct. TEVp expression was we

SUBMITTER: Williams DJ 

PROVIDER: S-EPMC2760398 | biostudies-literature | 2009 Oct

REPOSITORIES: biostudies-literature