Article
Conformation coupled enzyme catalysis: single-molecule and transient kinetics investigation of dihydrofolate reductase.
Biochemistry - 27 Dec 2005
Antikainen Nina M, Smiley R Derike, Benkovic Stephen J, Hammes Gordon G
Abstract excerpt
Ensemble kinetics and single-molecule fluorescence microscopy were used to study conformational transitions associated with enzyme catalysis by dihydrofolate reductase (DHFR). The active site loop of DHFR was labeled with a fluorescence quencher, QSY35, at amino acid position 17, and the fluorescent probe, Alexa555, at amino acid 37, by introducing cysteines at these sites with site-specific mutagenesis. The...
Topics
- Algorithms
- Biotinylation
- Catalysis
- Cysteine
- Escherichia coli
- Fluorescence Resonance Energy Transfer
- Fluorescent Dyes
- Folic Acid
- Hydrogen-Ion Concentration
- Kinetics
- Least-Squares Analysis
- Models, Chemical
- Mutagenesis, Site-Directed
- Mutation