Substrate profiling of tobacco etch virus protease using a novel fluorescence-assisted whole-cell assay.

ABSTRACT: Site-specific proteolysis of proteins plays an important role in many cellular functions and is often key to the virulence of infectious organisms. Efficient methods for characterization of proteases and their substrates will therefore help us understand these fundamental processes and thereby hopefully point towards new therapeutic strategies. Here, a novel whole-cell in vivo method was used to investigate the substrate preference of the sequence specific tobacco etch virus protease (TEVp). The assay, which utilizes protease-mediated intracellular rescue of genetically encoded short-lived fluorescent substrate reporters to enhance the fluorescence of the entire cell, allowed subtle differences in the processing efficiency of closely related substrate peptides to be detected. Quantitative

SUBMITTER: Kostallas G 

PROVIDER: S-EPMC3022733 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature