title: Comparative proteomics reveals novel components at the plasma membrane of differentiated HepaRG cells and different distribution in hepatocyte- and biliary-like cells.
abstract: Hepatitis B virus (HBV) is a human pathogen causing severe liver disease and eventually death. Despite important progress in deciphering HBV internalization, the early virus-cell interactions leading to infection are not known. HepaRG is a human bipotent liver cell line bearing the unique ability to differentiate towards a mixture of hepatocyte- and biliary-like cells. In addition to expressing metabolic functions normally found in liver, differentiated HepaRG cells support HBV infection in vitro, thus resembling cultured primary hepatocytes more than other hepatoma cells. Therefore, extensive characterization of the plasma membrane proteome from HepaRG cells would allow the identification of new cellular factors potentially involved in infection. Here we analyzed the plasma membranes of non-differentiated and differentiated HepaRG cells using nanoliquid chromatography-tandem mass spectrometry to identify the differences between the proteomes and the changes that lead to differentiation of these cells. We followed up on differentially-regulated proteins in hepatocytes- and biliary-like cells, focusing on Cathepsins D and K, Cyclophilin A, Annexin 1/A1, PDI and PDI A4/ERp72. Major differences between the two proteomes were found, including differentially regulated proteins, protein-protein interactions and intracellular localizations following differentiation. The results advance our current understanding of HepaRG differentiation and the unique properties of these cells.
authors: Department of Viral Glycoproteins, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania ; Department of Analytical Chemistry and Enviromental Engineering, Faculty of Applied Chemistry and Materials Science, Politehnica University of Bucharest, Bucharest, Romania.
authors: Catalina
authors: C
authors: Petrareanu
authors: Alina
authors: A
authors: Macovei
authors: Izabela
authors: I
authors: Sokolowska
authors: Alisa G
authors: AG
authors: Woods
authors: Catalin
authors: C
authors: Lazar
authors: Gabriel L
authors: GL
authors: Radu
authors: Costel C
authors: CC
authors: Darie
authors: Norica
authors: N
authors: Branza-Nichita
mesh_terms: D000595:Amino Acid Sequence; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D021381:Protein Transport; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics; D015854:Up-Regulation
chemical_list: D008565:Membrane Proteins; D010446:Peptide Fragments; D020543:Proteome
references: J Virol. 2010 Oct;84(19):9783-92
references: 20631122
references: Electrophoresis. 2012 Aug;33(16):2527-36
references: 22899260
references: J Cell Physiol. 2002 Nov;193(2):154-63
references: 12384992
references: Viruses. 2013 Jan 15;5(1):182-91
references: 23322171
references: J Pharmacol Exp Ther. 2012 Oct;343(1):134-44
references: 22776955
references: J Biol Chem. 2012 Jan 13;287(3):1719-33
references: 22130669
references: Eur J Biochem. 1995 Oct 1;233(1):1-8
references: 7588731
references: J Cell Biol. 1993 Jan;120(2):437-49
references: 8421057
references: Int J Biochem Cell Biol. 1999 Oct;31(10):1111-37
references: 10582342
references: Chem Biol Interact. 2007 May 20;168(1):66-73
references: 17241619
references: J Virol. 2013 Jun;87(11):6415-27
references: 23536683
references: J Proteome Res. 2009 Jan;8(1):118-22
references: 19053806
references: J Gen Virol. 2012 Mar;93(Pt 3):640-650
references: 22113009
references: Hepatology. 1996 Jul;24(1):1-5
references: 8707245
references: Proteomics. 2011 Dec;11(23):4514-28
references: 21932443
references: Methods Mol Biol. 2010;640:261-72
references: 20645056
references: J Mol Psychiatry. 2013 Jun 05;1(1):8
references: 25408901
references: Biol Chem. 1996 Nov;377(11):695-702
references: 8960370
references: J Virol. 2011 Nov;85(22):11809-20
references: 21900167
references: World J Gastroenterol. 2007 Jan 7;13(1):22-38
references: 17206752
references: Dev Dyn. 2001 Oct;222(2):206-17
references: 11668598
references: FEBS J. 2012 Jul;279(14):2579-94
references: 22613557
references: Nature. 2003 Mar 13;422(6928):198-207
references: 12634793
references: Cell Biol Toxicol. 2012 Apr;28(2):69-87
references: 22258563
references: Trends Microbiol. 2011 Apr;19(4):191-7
references: 21377881
references: Genome Biol. 2008 Jan 25;9(1):R19
references: 18221535
references: Hepatology. 2011 Aug;54(2):406-17
references: 21520209
references: Genomics. 2005 Dec;86(6):627-37
references: 16269234
references: Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15655-60
references: 12432097
references: Proteomics. 2013 Feb;13(3-4):538-57
references: 23193082
references: Hepatology. 1999 Feb;29(2):576-84
references: 9918938
references: J Virol. 2010 Jan;84(1):243-53
references: 19846513
references: Dev Biol. 2007 Oct 15;310(2):416-29
references: 17714699
references: J Biol Chem. 2001 May 11;276(19):15571-4
references: 11274138
references: Adv Genet. 2011;73:121-83
references: 21310296
references: J Virol. 2012 Feb;86(3):1789-801
references: 22114333
references: Lancet. 2001 Feb 17;357(9255):528-9
references: 11229675
references: Drug Metab Dispos. 2008 Jun;36(6):1111-8
references: 18347083
references: Proteomics. 2007 Jun;7(13):2201-15
references: 17549793
references: Nucleic Acids Res. 2011 Jan;39(Database issue):D561-8
references: 21045058
references: Methods Mol Biol. 2010;640:463-73
references: 20645068
references: J Virol. 2010 Apr;84(7):3373-81
references: 20089655
references: Biochem J. 1976 Apr 1;155(1):117-25
references: 938470
references: Nat Rev Microbiol. 2011 May;9(5):369-81
references: 21478902
references: J Mol Psychiatry. 2013 May 21;1(1):6
references: 25408899
references: Hepatology. 2007 Apr;45(4):957-67
references: 17393521
references: Electrophoresis. 1997 Mar-Apr;18(3-4):588-98
references: 9150946
references: Mol Cell Proteomics. 2005 Sep;4(9):1265-72
references: 15958392
references: Hepatology. 2012 Feb;55(2):373-83
references: 21953491
references: Mol Biol Cell. 2002 Dec;13(12):4456-69
references: 12475965
references: Biochem Biophys Res Commun. 2012 Mar 9;419(2):305-8
references: 22342715
references: Proteome Sci. 2012 Aug 02;10(1):47
references: 22857383
references: J Cell Biochem. 2012 May;113(5):1681-91
references: 22422622
references: Nucleic Acids Res. 2009 Jan;37(Database issue):D412-6
references: 18940858