hepatitis C virus;
NS5A protein;
conformational variation;
direct-acting antivirals;
drug resistance;
daclatasvir;
NONSTRUCTURAL PROTEIN 5A;
DIRECT-ACTING ANTIVIRALS;
CRYSTAL-STRUCTURE;
INHIBITOR BMS-790052;
IN-VITRO;
REPLICATION;
COMPLEX;
SCATTERING;
RESISTANCE;
INTERFERON;
D O I:
10.1099/jgv.0.001000
中图分类号:
Q81 [生物工程学(生物技术)];
Q93 [微生物学];
学科分类号:
071005 ;
0836 ;
090102 ;
100705 ;
摘要:
Direct-acting antivirals (DAAs) targeting the non-structural 5A (NS5A) protein of the hepatitis C virus (HCV) are crucial drugs that have shown exceptional clinical success in patients. However, their mode of action (MoA) remains unclear, and drug-resistant HCV strains are rapidly emerging. It is critical to characterize the behaviour of the NS5A protein in solution, which can facilitate the development of new classes of inhibitors or improve the efficacy of the currently available DAAs. Using biophysical methods, including dynamic light scattering, size exclusion chromatography and chemical cross-linking experiments, we showed that the NS5A domain 1 from genotypes 1b and 1a of the HCV intrinsically self-associated and existed as a heterogeneous mixture in solution. Interestingly, the NS5A domain 1 from genotypes 1 b and 1 a exhibited different dynamic equilibria of monomers to higher-order structures. Using small-angle X-ray scattering, we studied the structural dynamics of the various states of the NS5A domain 1 in solution. We also tested the effect of daclatasvir (DCV), the most prominent DAA, on self-association of the wild and DCV-resistant mutant (Y93H) NS5A domain 1 proteins, and demonstrated that DCV induced the formation of large and irreversible protein aggregates that eventually precipitated out. This study highlights the conformational variability of the NS5A domain 1 of HCV, which may be an intrinsic structural behaviour of the HCV NS5A domain 1 in solution.
机构:
Univ Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
Hop Beaujon, INSERM, U773, CRB3, F-92110 Clichy, FranceUniv Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
Asselah, Tarik
Marcellin, Patrick
论文数: 0引用数: 0
h-index: 0
机构:Univ Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
机构:
Univ Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
Fayoum Univ, Dept Engn Math & Phys, Al Fayyum, EgyptUniv Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
Barakat, Khaled H.
Anwar-Mohamed, Anwar
论文数: 0引用数: 0
h-index: 0
机构:
Univ Alberta, Li Ka Shing Inst Virol, Edmonton, AB, CanadaUniv Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Gaetano
Ardiri, Annalisa
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Ardiri, Annalisa
Proiti, Maria
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Proiti, Maria
Rigano, Giuseppe
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Rigano, Giuseppe
Frazzetto, Evelise
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Frazzetto, Evelise
Demma, Shirin
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Demma, Shirin
Ruggeri, Maria Irene
论文数: 0引用数: 0
h-index: 0
机构:
ARNAS Civ Hosp, Internal Med Unit, I-90142 Palermo, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Ruggeri, Maria Irene
Scuderi, Laura
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Scuderi, Laura
Malaguarnera, Giulia
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, Italy
Univ Catania, Dept Biomed Sci, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Malaguarnera, Giulia
Bertino, Nicoletta
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Fac Pharm, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Nicoletta
论文数: 引用数:
h-index:
机构:
Rapisarda, Venerando
Di Carlo, Isidoro
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Surg Sci Organ Transplantat & Adv Technol, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Di Carlo, Isidoro
Toro, Adriana
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Surg Sci Organ Transplantat & Adv Technol, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Toro, Adriana
Salomone, Federico
论文数: 0引用数: 0
h-index: 0
机构:
Acireale Hosp, Gastroenterol Unit, I-95024 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Salomone, Federico
Malaguarnera, Mariano
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Malaguarnera, Mariano
Bertino, Emanuele
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Fac Pharm, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Emanuele
Malaguarnera, Michele
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, Italy
Univ Catania, Dept Biomed Sci, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
机构:
Univ Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
Hop Beaujon, INSERM, U773, CRB3, F-92110 Clichy, FranceUniv Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
Asselah, Tarik
Marcellin, Patrick
论文数: 0引用数: 0
h-index: 0
机构:Univ Paris 07, Hepatol Dept, AP HP, F-92110 Clichy, France
机构:
Univ Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
Fayoum Univ, Dept Engn Math & Phys, Al Fayyum, EgyptUniv Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
Barakat, Khaled H.
Anwar-Mohamed, Anwar
论文数: 0引用数: 0
h-index: 0
机构:
Univ Alberta, Li Ka Shing Inst Virol, Edmonton, AB, CanadaUniv Alberta, Li Ka Shing Inst Virol, Edmonton, AB, Canada
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Gaetano
Ardiri, Annalisa
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Ardiri, Annalisa
Proiti, Maria
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Proiti, Maria
Rigano, Giuseppe
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Rigano, Giuseppe
Frazzetto, Evelise
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Frazzetto, Evelise
Demma, Shirin
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Demma, Shirin
Ruggeri, Maria Irene
论文数: 0引用数: 0
h-index: 0
机构:
ARNAS Civ Hosp, Internal Med Unit, I-90142 Palermo, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Ruggeri, Maria Irene
Scuderi, Laura
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Scuderi, Laura
Malaguarnera, Giulia
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, Italy
Univ Catania, Dept Biomed Sci, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Malaguarnera, Giulia
Bertino, Nicoletta
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Fac Pharm, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Nicoletta
论文数: 引用数:
h-index:
机构:
Rapisarda, Venerando
Di Carlo, Isidoro
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Surg Sci Organ Transplantat & Adv Technol, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Di Carlo, Isidoro
Toro, Adriana
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Dept Surg Sci Organ Transplantat & Adv Technol, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Toro, Adriana
Salomone, Federico
论文数: 0引用数: 0
h-index: 0
机构:
Acireale Hosp, Gastroenterol Unit, I-95024 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Salomone, Federico
Malaguarnera, Mariano
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Malaguarnera, Mariano
Bertino, Emanuele
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Fac Pharm, I-95123 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy
Bertino, Emanuele
Malaguarnera, Michele
论文数: 0引用数: 0
h-index: 0
机构:
Univ Catania, Res Ctr La Grande Senesce, I-95100 Catania, Italy
Univ Catania, Dept Biomed Sci, I-95100 Catania, ItalyUniv Catania, Dept Clin & Expt Med, Hepatol Unit, I-95123 Catania, Italy