An investigation into the minimum requirements for peptide hydrolysis by mutation of the catalytic triad of trypsin

摘要: The catalytic triad of rat anionic trypsin has been systematically altered by site-directed mutagenesis to determine the activity alternate combinations amino acids toward hydrolysis peptide bonds. Genetically modified trypsins H57A, H57D, H57E, H57K, H57R, H57A/D102N, H57D/D102N, H57L/D102N, H57K/D102N, D102JN, S195A, S195T and H57A/D102N/S195A have generated. Rigorous steps were taken show that resultant catalysis was due mutant enzymes not contaminants