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 > 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.
Varadarajan
Navin
N
Institute for Cell and Molecular Biology, University of Texas, Austin, Texas 78712, USA.
Rodriguez
Sarah
S
Hwang
Bum-Yeol
BY
Georgiou
George
G
Iverson
Brent L
BL
D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D007700:Kinetics; D015202:Protein Engineering; D013379:Substrate Specificity; D014633:Valine / Q000378:metabolism
D010450:Endopeptidases; D014633:Valine