Highly active and selective endopeptidases with programmed substrate specificities


A family of engineered endopeptidases has been created that is capable of cleaving a diverse array of peptide sequences with high selectivity and catalytic efficiency (kcat/KM > 104 M?1 s?1). By screening libraries with a selection-counterselection substrate method, protease variants were programmed to recognize amino acids having altered charge, size and hydrophobicity properties adjacent to the scissile bond of the substrate, including Glu?Arg, a specificity that to our knowledge has not been observed among natural proteases. Members of this artificial protease family resulted from a relatively small number of amino acid substitutions that (at least in one case) proved to be epistatic.




Copyright 2008 Nature Chemical Biology. This is post-print version of a published paper that is available at: https://www.nature.com/articles/nchembio.80. Recommended citation: Varadarajan, Navin, Sarah Rodriguez, Bum-Yeol Hwang, George Georgiou, and Brent L. Iverson. "Highly active and selective endopeptidases with programmed substrate specificities." Nature chemical biology 4, no. 5 (2008): 290. doi: 10.1038/nchembio.80. This item has been deposited in accordance with the publisher copyright and licensing terms and with the author's permission.