Work

Directed Evolution of Highly Selective Proteases by Using a Novel FACS‐Based Screen that Capitalizes on the p53 Regulator MDM2

Year: 2012

Type: article

Abstract: We describe a novel method for the isolation of selective protease variants displayed on the surface of E. coli. The method relies on the electrostatic capture of an autoinhibited protein on the cell surface, combined with external labeling using a fluorophore-conjugated binding peptide (PMI-FL). Using this method, we isolated an OmpT variant that hydrolyzes a target sequence (between Ala-Arg) with high selectivity.

Source: ChemBioChem

DOI: 10.1002/cbic.201100718

Authors: Tae Hyeon Yoo, Mark Pogson, Brent L. Iverson, George Georgiou

Institutions: Ajou University, The University of Texas at Austin

Language: English

Topic: Enzyme Catalysis and Immobilization

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, National Institute of General Medical Sciences

Awards: Protease Engineering, G8527580684