Article
A protein engineering study of the role of aspartate 158 in the catalytic mechanism of papain.
Biochemistry - 17 Jul 1990
Ménard R, Khouri H E, Plouffe C, Dupras R, Ripoll D, Vernet T, Tessier D C, Lalberté F, Thomas D Y, Storer A C
Abstract excerpt
The controversy concerning the various suggested roles for the side chain of Asp158 in the active site of papain has been clarified by using site-directed mutagenesis. Both wild-type papain and an Asp158 Asn variant were produced in a baculovirus-insect cell expression system, purified to homogeneity from the culture, and characterized kinetically. With CBZ-Phe-Arg-MCA as substrate, the kcat/KM and kcat values...
Topics
- Animals
- Aspartic Acid
- Base Sequence
- Cells, Cultured
- Enzyme Induction
- Genes, Synthetic
- Hydrogen-Ion Concentration
- Insecta
- Kinetics
- Molecular Sequence Data
- Mutation