ABSTRACT
Comparative study of DNA biosynthesis inhibition, catalyzed by avian myeloblastose virus (AMV) reverse transcriptase (RT), human immunodeficiency virus (HIV) recombinant and native RT, has been performed. 3'-Azido-2',3'-dideoxythymidine 5'-triphosphate (AzTTP); 3'-azido-2',3'-dideoxythymidine 5'-methylenephosphonate-diphosphate: 3'-azido-2',3'-dideoxythymidine 5'-phosphate-phosponoacetate; 3'-azido-2',3'-dideoxythymidine 5'-phosphate-dibromomethylenephosphonate; 2',3'-O-isopropylidenecytidine 5'-methylenephosphonate-diphosphate (rC-IP-MPDP) were used as inhibitors. AzTTP proved to by the most active inhibitor (its activity against HIV RT is higher than against AMV RT), although not selective as the phosphonates; only rC-iP-MPDP has low selectivity.
Subject(s)
DNA/biosynthesis , Retroviridae/enzymology , Reverse Transcriptase Inhibitors , Avian Myeloblastosis Virus/enzymology , Avian Sarcoma Viruses/enzymology , Catalysis , Chemical Phenomena , Chemistry , HIV/enzymology , Oligodeoxyribonucleotides/metabolism , Poly A , Templates, Genetic , Zidovudine/analogs & derivatives , Zidovudine/pharmacologyABSTRACT
A hybrid plasmid pPR6 was constructed containing BgII-EcoRI fragment of the pol region of HIV (strain IIIB) genome which determined the synthesis of virus-specific protease. Extracts of E. coli DN5/pPR6 bacteria provided for specific hydrolysis of hybrid protein p165 (the N-terminus of which is presented by complete beta-galactosidase and the C-terminus by duplicated area of virus-specific precursor p55 containing a site for virus-specific protease located at the border of proteins p17 and p24) with formation of products having molecular weights of 19, 42, 28, 23, and 19 kD. Polypeptides 119K, 23K, and 19K are products of complete hydrolysis, and 42K a result of partial cleavage. The kinetics of hydrolysis in relation to pH values of the reaction mixture was analysed. It is suggested that the reported system of HIV protease activity determination be used for screening of potential inhibitors of this enzyme.
Subject(s)
Escherichia coli/enzymology , HIV Protease/analysis , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Gene Expression Regulation, Enzymologic/genetics , Gene Expression Regulation, Viral/genetics , Genes, Viral/genetics , HIV Protease/biosynthesis , HIV Protease/genetics , HIV-1/enzymology , HIV-1/genetics , Hydrogen-Ion Concentration , Hydrolysis , Immunoblotting , Molecular Sequence Data , Molecular Weight , Plasmids/genetics , Recombination, GeneticABSTRACT
E. coli cells growing on the medium containing glucose and lactate do not utilize lactate. One reason of preferential utilization of glucose is catabolite inhibition of lactate transport. It is necessary for glucose to penetrate into the cell to inhibit lactate transport. Besides glucose the inhibition of the lactate transport is also caused by fructose and by non-metabolized analogue of glucose--alpha-methylglucoside.