Recombinant poliovirus 3C protease. The enzyme application to protein specific fragmentation.
Active 3C protease of poliovirus 1(M) was obtained when cloning and expressing fragment HindII-HindIII (bases from 5240 to 6056) of cDNA in vector pTTQ8 in E.coli cells. As shown, fragment 3D of polymerase covalently bound to 3C does not deprive the enzyme of its specific proteolytic activity. The absence of 26 N-terminal amino acids in 3C entails its inactivation. The recombinant 3C protease cleaved peptide bond Gln-Gly not only in virus polyprotein, but also in molecules of beta-galactosidase and bovine catalase.
Sablina
E P
EP
Shemyakin Institute of Bioogranic Chemistry, USSR Academy of Sciences, Moscow.
Antonov
V K
VK
D000086042:3C Viral Proteases; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D007460:Iodoacetamide / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017955:Poliovirus / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D013379:Substrate Specificity; D014764:Viral Proteins; D001616:beta-Galactosidase / Q000378:metabolism
D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D002374:Catalase; D001616:beta-Galactosidase; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases; D007460:Iodoacetamide