کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6134256 | 1593482 | 2013 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Humanized-VH/VHH that inhibit HCV replication by interfering with the virus helicase activity
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کلمات کلیدی
RT-PCRVHHcDNA - cDNAVirus neutralization assay - آزمایش خنثی سازی ویروسcomplementary deoxyribonucleic acid - اسید دز اکسید ریبونوکلئیک مکملRNA - اسید ریبونوکلئیکribonucleic acid - اسید ریبونوکلئیکFluorescent Resonance Energy Transfer - انتقال انرژی رزونانس فلورسنتFRET - انتقال انرژی رزونانسی فورسترEpitope - اپیتوپHCV - هپاتیت سیHepatitis C virus - هپاتیت سیpolymerase chain reaction - واکنش زنجیره ای پلیمرازreverse transcription polymerase chain reaction - واکنش زنجیره ای پلیمراز رونویسی معکوسPCR - واکنش زنجیرهٔ پلیمرازoptical density - چگالی نوری
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
NS3 helicase is a pivotal enzyme involved in the early and late phases of hepatitis C virus (HCV) replication. The primary sequence and tertiary structure of this virus enzyme differ from human helicase to a certain extent; thus this virus protein has potential as a novel anti-HCV target. In this study, recombinant C-terminal NS3 protein of HCV genotype 3a with endowed helicase activity was produced and used as antigen by selecting VH/VHH display phage clones from an established humanized-camel single domain antibody library that bound specifically to HCV helicase. The VH/VHH derived from phage transfected Escherichia coli clones were linked molecularly to a cell penetrating peptide, i.e., penetratin (PEN). The cell penetrable VH/VHH (transbodies) could reduce the amounts of the HCV RNA released into the cell culture fluid and inside Huh7 cells infected with pJFH1 replicon with a greater effect on the former compared to the latter. Regions and residues of the helicase bound by the transbodies were determined by phage mimotope searching and multiple alignments as well as homology modeling and molecular docking. The epitope of one transbody (PEN-VHH9) encompassed residues 588RLKPTLHGPTPLLYRLGA605 of the domain 3 necessary for helicase activity while another transbody (PEN-VH59) interacted with the areas covering the phenylalanine loop and arginine clamp of the domain 2 which are important for the proper folding of the enzyme as well as nucleic acid substrate binding. Although the molecular mechanisms of the prototypic transbodies on NS3 helicase need further investigation, these transbodies have high potential as novel, safe and mutation tolerable anti-HCV agents.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 194, Issues 1â2, December 2013, Pages 289-299
Journal: Journal of Virological Methods - Volume 194, Issues 1â2, December 2013, Pages 289-299
نویسندگان
Aninthita Phalaphol, Kanyarat Thueng-in, Jeeraphong Thanongsaksrikul, Ornnuthchar Poungpair, Kunan Bangphoomi, Nitat Sookrung, Potjanee Srimanote, Wanpen Chaicumpa,