title: Identification of temperature-sensitive mutants of the human immunodeficiency virus type 1 protease through saturation mutagenesis. Amino acid side chain requirements for temperature sensitivity.
abstract: Human immunodeficiency virus type 1 encodes a protease whose activity is required for the production of infectious virus. An Escherichia coli expression and processing assay system was used to screen 285 protease mutants for temperature-sensitive activity. Fourteen protease mutants had a temperature-sensitive phenotype, and approximately half resulted from conservative amino acid substitutions. Of the 14 substitutions that conferred a temperature-sensitive phenotype, 11 substitutions occurred at 6 positions that represent 3 pairs of residues in the protease that contact each other in the three-dimensional structure. These mutants assist in pinpointing regions of the protease that are important for enzyme activity and stability.
authors.affiliation: University of North Carolina, Lineberger Comprehensive Cancer Center, Chapel Hill 27599.
authors.forename: M
authors.initials: M
authors.lastname: Manchester
authors.forename: L
authors.initials: L
authors.lastname: Everitt
authors.forename: D D
authors.initials: DD
authors.lastname: Loeb
authors.forename: C A
authors.initials: CA
authors.lastname: Hutchison
authors.forename: R
authors.initials: R
authors.lastname: Swanstrom
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D004795:Enzyme Stability; D016333:HIV Protease / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D010641:Phenotype; D011487:Protein Conformation; D013696:Temperature
chemical_list: D000596:Amino Acids; D016333:HIV Protease