Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus
Research output: Contribution to journal › Article
Transient kinetics of the equine infectious anemia virus deoxyuridine 5'-triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one-step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis.
|Research areas and keywords||
Subject classification (UKÄ) – MANDATORY
|Publication status||Published - 2000|
Related research output
Nucleotide Binding to Trimeric dUTPase: Studies on Recombinant Enzymes using Kinetic and Spectroscopic MethodsNord, J., 2000, Johan Nord, Department of Biochemistry, Center for Chemistry and Chemical Engineering, P.O. Box 124, S-221 00, Lund, Sweden. 142 p.
Research output: Thesis › Doctoral Thesis (compilation)