title: Highly active and selective endopeptidases with programmed substrate specificities.
abstract: 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 > 10(40 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 GluArg, 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.
authors.affiliation: Institute for Cell and Molecular Biology, University of Texas, Austin, Texas 78712, USA.
authors.forename: Navin
authors.initials: N
authors.lastname: Varadarajan
authors.forename: Sarah
authors.initials: S
authors.lastname: Rodriguez
authors.forename: Bum-Yeol
authors.initials: BY
authors.lastname: Hwang
authors.forename: George
authors.initials: G
authors.lastname: Georgiou
authors.forename: Brent L
authors.initials: BL
authors.lastname: Iverson
mesh_terms: D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D007700:Kinetics; D015202:Protein Engineering; D013379:Substrate Specificity; D014633:Valine / Q000378:metabolism
chemical_list: D010450:Endopeptidases; D014633:Valine