title: An engineered retroviral proteinase from myeloblastosis associated virus acquires pH dependence and substrate specificity of the HIV-1 proteinase.
abstract: In an attempt to understand the structural reasons for differences in specificity and activity of proteinases from two retroviruses encoded by human immunodeficiency virus (HIV) and myeloblastosis associated virus (MAV), we mutated five key residues predicted to form part of the enzyme subsites S1, S2 and S3 in the substrate binding cleft of the wild-type MAV proteinase wMAV PR. These were changed to the residues occupying a similar or identical position in the HIV-1 enzyme. The resultant mutated MAV proteinase (mMAV PR) exhibits increased enzymatic activity, altered substrate specificity, a substantially changed pH activity profile and a higher pH stability close to that observed in the HIV-1 PR. This dramatic alteration of MAV PR activity achieved by site-directed mutagenesis suggests that we have identified the amino acid residues contributing substantially to the differences between MAV and HIV-1 proteinases.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, Praha.
authors.forename: J
authors.initials: J
authors.lastname: Konvalinka
authors.forename: M
authors.initials: M
authors.lastname: Horejsí
authors.forename: M
authors.initials: M
authors.lastname: Andreánsky
authors.forename: P
authors.initials: P
authors.lastname: Novek
authors.forename: I
authors.initials: I
authors.lastname: Pichová
authors.forename: I
authors.initials: I
authors.lastname: Bláha
authors.forename: M
authors.initials: M
authors.lastname: Fábry
authors.forename: J
authors.initials: J
authors.lastname: Sedlácek
authors.forename: S
authors.initials: S
authors.lastname: Foundling
authors.forename: P
authors.initials: P
authors.lastname: Strop
mesh_terms: D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D016333:HIV Protease / Q000235:genetics* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D015202:Protein Engineering; D012190:Retroviridae / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D010450:Endopeptidases; D016333:HIV Protease