دورية أكاديمية
Co-evolution of drug resistance and broadened substrate recognition in HIV protease variants isolated from an Escherichia coli genetic selection system.
| العنوان: | Co-evolution of drug resistance and broadened substrate recognition in HIV protease variants isolated from an Escherichia coli genetic selection system. |
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| المؤلفون: | Koivisto JM; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Poulsen NR; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Larsen BS; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Weibull MGM; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Stein A; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Doro F; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Winther JR; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Lindorff-Larsen K; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark., Willemoës M; From the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200N, Denmark. |
| المصدر: | The Biochemical journal [Biochem J] 2022 Feb 17; Vol. 479 (4), pp. 479-501. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Published by Portland Press on behalf of the Biochemical Society Country of Publication: England NLM ID: 2984726R Publication Model: Print Cited Medium: Internet ISSN: 1470-8728 (Electronic) Linking ISSN: 02646021 NLM ISO Abbreviation: Biochem J Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: London, UK : Published by Portland Press on behalf of the Biochemical Society |
| مواضيع طبية MeSH: | Anti-HIV Agents/*pharmacokinetics , Drug Resistance, Viral/*genetics , HIV Protease/*genetics, Amino Acid Substitution ; AraC Transcription Factor/genetics ; Arabinose/metabolism ; Chymosin/metabolism ; Escherichia coli ; Escherichia coli Proteins/genetics ; Fusion Proteins, gag-pol/metabolism ; Gene Products, gag/metabolism ; Genes, araC ; HIV Protease/chemistry ; HIV Protease/isolation & purification ; HIV Protease/metabolism ; Models, Molecular ; Mutation, Missense ; Point Mutation ; Protein Conformation ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Saquinavir/antagonists & inhibitors ; Saquinavir/pharmacology ; Selection, Genetic ; Sequence Alignment ; Sequence Homology, Amino Acid ; Structure-Activity Relationship ; Substrate Specificity |
| مستخلص: | A genetic selection system for activity of HIV protease is described that is based on a synthetic substrate constructed as a modified AraC regulatory protein that when cleaved stimulate l-arabinose metabolism in an Escherichia coli araC strain. Growth stimulation on selective plates was shown to depend on active HIV protease and the scissile bond in the substrate. In addition, the growth of cells correlated well with the established cleavage efficiency of the sites in the viral polyprotein, Gag, when these sites were individually introduced into the synthetic substrate of the selection system. Plasmids encoding protease variants selected based on stimulation of cell growth in the presence of saquinavir or cleavage of a site not cleaved by wild-type protease, were indistinguishable with respect to both phenotypes. Also, both groups of selected plasmids encoded side chain substitutions known from clinical isolates or displayed different side chain substitutions but at identical positions. One highly frequent side chain substitution, E34V, not regarded as a major drug resistance substitution was found in variants obtained under both selective conditions and is suggested to improve protease processing of the synthetic substrate. This substitution is away from the substrate-binding cavity and together with other substitutions in the selected reading frames supports the previous suggestion of a substrate-binding site extended from the active site binding pocket itself. (© 2022 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.) |
| فهرسة مساهمة: | Keywords: E coli genetic selection system; AraC protein; random mutagenesis analysis; retro viral protease; substrate recognition |
| المشرفين على المادة: | 0 (Anti-HIV Agents) 0 (AraC Transcription Factor) 0 (AraC protein, E coli) 0 (Escherichia coli Proteins) 0 (Fusion Proteins, gag-pol) 0 (Gene Products, gag) 0 (Recombinant Fusion Proteins) B40ROO395Z (Arabinose) EC 3.4.23.- (HIV Protease) EC 3.4.23.- (p16 protease, Human immunodeficiency virus 1) EC 3.4.23.4 (Chymosin) L3JE09KZ2F (Saquinavir) |
| تواريخ الأحداث: | Date Created: 20220128 Date Completed: 20220228 Latest Revision: 20220228 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1042/BCJ20210767 |
| PMID: | 35089310 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1470-8728 |
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| DOI: | 10.1042/BCJ20210767 |