Reconstitution of hepatitis C virus envelope glycoproteins into liposomes as a surrogate model to study virus attachment.
The envelope glycoproteins, E1 and E2, of hepatitis C virus (HCV) assemble intracellularly to form a noncovalent heterodimer that is expected to be essential for viral assembly and entry. However, due to the lack of a cell culture system supporting efficient HCV replication, it is very difficult to obtain relevant information on the functions of this glycoprotein oligomer. To get better insights into its biological and biochemical properties, HCV envelope glycoprotein heterodimer expressed by a vaccinia virus recombinant was purified by immunoaffinity. Purified E1E2 heterodimer was recognized by conformation-dependent monoclonal antibodies, showing that the proteins were properly folded. In addition, it interacted with human CD81, a putative HCV receptor, as well as with human low and very low density lipoproteins, which have been shown to be associated with infectious HCV particles isolated from patients. Purified E1E2 heterodimer was also reconstituted into liposomes. E1E2-liposomes were recognized by a conformation-dependent monoclonal antibody as well as by human CD81. Together, these data indicate that E1E2-liposomes are a valuable tool to study the molecular requirements for HCV binding to target cells.
Lambot
Michel
M
CNRS-Institut de Biologie de Lille & Institut Pasteur de Lille, 59021 Lille Cedex, France.
Frétier
Stéphanie
S
Op De Beeck
Anne
A
Quatannens
Brigitte
B
Lestavel
Sophie
S
Clavey
Veronique
V
Dubuisson
Jean
J
D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D016174:Hepacivirus / Q000502:physiology*; D008081:Liposomes; D008561:Membrane Fusion; D014759:Viral Envelope Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000302:isolation & purification / Q000378:metabolism*
D008081:Liposomes; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C107842:protein E1, Classical swine fever virus; C091015:glycoprotein E2, Hepatitis C virus