title: Specificity studies on retroviral proteinase from myeloblastosis-associated virus.
abstract: The specificity of the p15 proteinase of myeloblastosis-associated virus (MAV) was tested with nonviral high molecular weight substrates and with synthetic peptides. Peptides with sequences spanning known cleavage sites in viral polyproteins of Rous sarcoma virus (RSV) and avian leukemia viruses, as well as in BSA and HSA, were synthesized, and the rate of their cleavage by the MAV proteinase was compared. Synthetic peptides require for successful cleavage at least 4 residues at the N-terminal side and 3 residues at the C-terminal side. The proteinase shows a preference for hydrophobic residues with bulky side chains (Met, Tyr, Phe) in P3, although Arg and Gln can also be accepted. Small hydrophobic residues are required in P2 and P2', and large hydrophobic residues (Tyr, Met, Phe/p-nitro-Phe) are preferred in both P1 and P1'. The difference between the specificity of the p15 proteinase and that of the HIV-1 proteinase mostly pertains to position P2' of the substrate, where bulkier side chains are accepted by the HIV-1 proteinase (Richards et al., 1990). A good chromogenic substrate for the MAV and RSV proteinases was developed and used to further characterize the MAV proteinase activity with respect to ionic strength and pH. The activity of the proteinase is strongly dependent on ionic strength and pH. Both the kcat and Km values contribute to a higher cleavage efficiency at higher salt concentrations and show a bell-shaped pH dependence curve with a sharp maximum at pH 5.5 (kcat) and 6.5 (Km).
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Science, Prague.
authors.forename: P
authors.initials: P
authors.lastname: Strop
authors.forename: J
authors.initials: J
authors.lastname: Konvalinka
authors.forename: D
authors.initials: D
authors.lastname: Stys
authors.forename: L
authors.initials: L
authors.lastname: Pavlickova
authors.forename: I
authors.initials: I
authors.lastname: Blaha
authors.forename: J
authors.initials: J
authors.lastname: Velek
authors.forename: M
authors.initials: M
authors.lastname: Travnicek
authors.forename: V
authors.initials: V
authors.lastname: Kostka
authors.forename: J
authors.initials: J
authors.lastname: Sedlacek
mesh_terms: D000595:Amino Acid Sequence; D016282:Aspartic Acid Endopeptidases; D001354:Avian Leukosis Virus / Q000235:genetics; D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001665:Binding Sites; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D015750:Genes, gag; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D012191:Retroviridae Proteins / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D012191:Retroviridae Proteins; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D010450:Endopeptidases; D010447:Peptide Hydrolases; D016282:Aspartic Acid Endopeptidases; C036946:protease p15