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Proteomic analysis of plasma membranes isolated from undifferentiated and differentiated HepaRG cells

Izabela Sokolowska1, Cristina Dorobantu2, Alisa G Woods1, Alina Macovei2, Norica Branza-Nichita2* and Costel C Darie1*

Author Affiliations

1 Biochemistry & Proteomics Group, Department of Chemistry & Biomolecular Science, Clarkson University, 8 Clarkson Avenue, Potsdam, NY, 13699-5810, USA

2 Department of Glycoproteins, Institute of Biochemistry of the Romanian Academy, Splaiul Independentei, 296, Sector 6, Bucharest, 060031, Romania

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Proteome Science 2012, 10:47  doi:10.1186/1477-5956-10-47

Published: 2 August 2012


Liver infection with hepatitis B virus (HBV), a DNA virus of the Hepadnaviridae family, leads to severe disease, such as fibrosis, cirrhosis and hepatocellular carcinoma. The early steps of the viral life cycle are largely obscure and the host cell plasma membrane receptors are not known. HepaRG is the only proliferating cell line supporting HBV infection in vitro, following specific differentiation, allowing for investigation of new host host-cell factors involved in viral entry, within a more robust and reproducible environment. Viral infection generally begins with receptor recognition at the host cell surface, following highly specific cell-virus interactions. Most of these interactions are expected to take place at the plasma membrane of the HepaRG cells. In the present study, we used this cell line to explore changes between the plasma membrane of undifferentiated (−) and differentiated (+) cells and to identify differentially-regulated proteins or signaling networks that might potentially be involved in HBV entry. Our initial study identified a series of proteins that are differentially expressed in the plasma membrane of (−) and (+) cells and are good candidates for potential cell-virus interactions. To our knowledge, this is the first study using functional proteomics to study plasma membrane proteins from HepaRG cells, providing a platform for future experiments that will allow us to understand the cell-virus interaction and mechanism of HBV viral infection.

Hepatocytes; HBV; Proteomics; Mass spectrometry; Differentiation