کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020433 1542337 2014 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Production and characterization of the ectodomain of E2 envelope glycoprotein of hepatitis C virus folded in the presence of full-length E1 glycoprotein
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Production and characterization of the ectodomain of E2 envelope glycoprotein of hepatitis C virus folded in the presence of full-length E1 glycoprotein
چکیده انگلیسی


• The ectodomain of E2 was folded in a native conformation in the presence of E1.
• Its properties are the same than those of the ectodomain folded in the absence of E1.
• E2 ectodomain behaves as an independent folding unit.

Hepatitis C virus (HCV) envelope glycoproteins, E1 and E2, are involved in the first steps of virus infection. The E2 ectodomain can be produced as an isolated form (E2661). However, there is some concern about its proper conformation and the role that E1 can play as a chaperone for the folding of E2. In order to verify this fact we have expressed a chimeric protein (E1tmbE2) based on the full-length E1 sequence followed by the E2 ectodomain using the baculovirus-insect cells system. The E2 ectodomain is folded in the presence of the E1, proteolytically processed by cellular proteases and secreted to cell culture media (E2661p), while the E1 protein is retained into the cell due to its transmembrane sequence. The purification of E2661p from culture media was facilitated by a His tag introduced in its amino terminus. Both E2661 and E2661p glycoproteins shared very similar structural features, monitored by spectroscopic and antigenic studies. Moreover, their functional properties, tested by means of CD81 binding, were almost indistinguishable, indicating that the E2 ectodomain constitutes an independent folding unit.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 104, December 2014, Pages 20–25
نویسندگان
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