title: Identification of histone deacetylase 1 protein complexes in liver cancer cells.
abstract: BACKGROUND
  Hepatocellular carcinoma is one of the leading causes of mortalities worldwide. The search for new therapeutic targets is of utmost importance for improved treatment. Altered expression of HDAC1 in hepatocellular carcinoma (HCC) and its requirement for liver formation in zebrafish, suggest that it may regulate key events in liver carcinogenesis and organogenesis. However, molecular mechanisms of HDAC1 action in liver carcinogenesis are largely unknown. The present study was conducted to identify HDAC1 interacting proteins in HepG2 cells using modified SH-double-affinity purification coupled with liquid mass spectrophotemetery.
  
  
  METHODS
  HepG2 cells were transfected with a construct containing HDAC1 with a C-terminal strepIII-HA tag as bait. Bait proteins were confirmed to be expressed in HepG2 cells by western blotting and purified by double affinity columns and protein complexes for analysis on a Thermo LTQ Orbitrap XL using a C18 nano flow ESI liquid chromatography system.
  
  
  RESULTS
  There were 27 proteins which showed novel interactions with HDAC1 identified only in this study, while 14 were among the established interactors. Various subunits of T complex proteins (TCP1) and prefoldin proteins (PFDN) were identified as interacting partners that showed high affinity with HDAC1 in HepG2 cells.
  
  
  CONCLUSIONS
  The double affinity purification method adopted in this study was very successful in terms of specificity and reproducibility. The novel HDAC1 complex identified in this study could be better therapeutic target for treatment of hepatocellular carcinoma.
authors.affiliation: Bioproducts Research Group, Department of Zoology, College of Science, King Saud University, Riyadh, Kingdom of Saudi Arabia. fmuhammad@ksu.edu.sa
authors.forename: Muhammad
authors.initials: M
authors.lastname: Farooq
authors.forename: Wael N
authors.initials: WN
authors.lastname: Hozzein
authors.forename: Elsayed A
authors.initials: EA
authors.lastname: Elsayed
authors.forename: Nael A
authors.initials: NA
authors.lastname: Taha
authors.forename: Mohammad Am
authors.initials: MA
authors.lastname: Wadaan
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D056404:Chaperonin Containing TCP-1 / Q000378:metabolism*; D015870:Gene Expression; D056945:Hep G2 Cells; D056284:Histone Deacetylase 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D018832:Molecular Chaperones / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D018832:Molecular Chaperones; C533304:TCP1 protein, human; C112806:prefoldin; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D056404:Chaperonin Containing TCP-1