Hepatitis C virus: The role of N-glycosylation sites of viral genotype 1b proteins for formation of viral particles in insect and mammalian cells
| Autor: | Sergey N. Kochetkov, Drutsa Vl, Pavel Spirin, N. N. Koroleva, V.A. Orlov, V.S. Prassolov, Olga V. Orlova, S. N. Beljelarskaya, A.V. Timohova, Alexander V. Ivanov, Vladimir I. Popenko, Petr M. Rubtsov |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Glycosylation Hepatitis C virus Biophysics N-glycans Biology medicine.disease_cause Biochemistry Virus 03 medical and health sciences chemistry.chemical_compound N-linked glycosylation Genotype medicine HCV envelope glycoproteins Baculovirus chemistry.chemical_classification Site-directed mutagenesis Mammalian Hek293Tcells virus diseases RNA Virology Molecular biology NS2-3 protease 030104 developmental biology chemistry Insect Sf9 cells Glycoprotein Research Article |
| Zdroj: | Biochemistry and Biophysics Reports |
| ISSN: | 2405-5808 |
| DOI: | 10.1016/j.bbrep.2016.05.019 |
| Popis: | Hepatitis C virus (HCV) is characterized by considerable genetic variability and, as a consequence, it has 6 genotypes and multitude of subtypes. HCV envelope glycoproteins are involved in the virion formation; the correct folding of these proteins plays the key role in virus infectivity. Glycosylation at certain sites of different genotypes HCV glycoproteins shows substantial differences in functions of the individual glycans (Goffard et al., 2005; Helle et al., 2010) [1], [2]. In this study, differential glycosylation sites of HCV genotype 1b envelope proteins in insect and mammalian cells was demonstrated. We showed that part of glycosylation sites was important for folding of the proteins involved in the formation of viral particles. Point mutations were introduced in the protein N-glycosylation sites of HCV (genotype 1b) and the mutant proteins were analyzed using baculovirus expression system in mammalian and insect cells. Our data showed that, in contrast to HCV 1a and 2a, the folding of HCV 1b envelope proteins E2 (sites N1, N2, N10) and E1 (sites N1, N5) was disrupted, however that did not prevent the formation of virus-like particles (VLP) with misfolded glycoproteins having densities typical for HCV particles containing RNA fragments. Experimental data are supported by mathematical modeling of the structure of E1 mutant variants. Highlights • Baculovirus expression system in mammalian and insect cells for. • Glycosylation at certain sites of HCV genotype 1b. • Part of glycosylation sites of E1, E2 HCV important for folding of the proteins involved in the formation of viral particles. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|