Electrophoretic and spectral characterization of wild type and mutant adenovirus protease.
The P137L mutation in the adenovirus type 2 protease results in a temperature-sensitive protein-trafficking phenotype expressed during infection but not in vitro. Homology-derived secondary structure prediction placed the mutation within an externally disposed loop. Circular dichroism and urea gradient gel electrophoresis suggested that, unlike other thiol proteases, the Ad2 protease is comprised of a single conformational domain. The -0.32-kcal difference in the free energy of folding and the temperature-independent CD spectra of the mutant and wild type enzymes point to a very subtle structural change as the cause of the in vivo phenotype.
Keyvani-Amineh
H
H
Department of Microbiology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
Diouri
M
M
Guillemette
J G
JG
Weber
J M
JM
D000260:Adenoviruses, Human / Q000201:enzymology* / Q000235:genetics*; D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004586:Electrophoresis; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics*; D004795:Enzyme Stability; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010641:Phenotype; D017354:Point Mutation; D017433:Protein Structure, Secondary
D010450:Endopeptidases