Article
Dissecting substrate recognition by thrombin using the inactive mutant S195A.
Biophysical chemistry - 1 Jan 2003
Krem Maxwell M, Di Cera Enrico
Abstract excerpt
The catalytically inactive mutant S195A was used to study the interaction of thrombin with substrates under equilibrium conditions. By monitoring changes in intrinsic fluorescence, we measured dissociation constants for a variety of synthetic substrates, PAR peptides and the inhibitor PPACK. The S195A mutant retains the Na(+)-binding properties of the wild type, and substrate binding to the mutant is enhanced by...
Topics
- Algorithms
- Amino Acid Chloromethyl Ketones
- Chemical Phenomena
- Chemistry, Physical
- Fluorescent Dyes
- Humans
- Kinetics
- Ligands
- Mutagenesis, Site-Directed
- Mutation
- Spectrometry, Fluorescence
- Substrate Specificity
- Thrombin