title: Targeting Mechanism of a Novel Liver-targeting Interferon IFN-CSP Involves Liver Heparan Sulfate Proteoglycan.
abstract: BACKGROUND
  In our previous study, a novel liver-targeting interferon (IFN-CSP) combining IFN α2b with plasmodium region I-plus peptide was successfully designed and prepared with Escherichia coli expression systems. The purified IFN-CSP showed anti-HBV activity and liver-targeting potentiality. The present investigation was designed to investigate the molecular mechanisms responsible for liver-targeting of IFN-CSP.
  
  
  METHODS
  The binding site of IFN-CSP in hepatocytes was assayed by immunofluorescent staining. The correspondence of HSPG distribution and the pattern of IFN-CSP binding in liver tissue were determined using a confocal laser scanning microscope. Both the hepatocytes and liver tissue were using as model to investigate the effect of enzyme and soluble glycosaminoglycan on IFN-CSP binding using flow cytometry and fluorescence microscope.
  
  
  RESULTS
  Studies of hepatocytes demonstrated that the localization of IFN-CSP in hepatocytes was the plasma membrane. Studies of liver tissue slices showed that IFN-CSP bound to liver tissue in a pattern similar to the distribution of heparan sulfate proteoglycan (HSPG) immunoreactivity. Pretreatment of hepatocytes and liver slices with heparinase reduced the binding of IFN-CSP to HepG2.2.15 cells and liver slices. Coincubation of IFN-CSP with heparin markedly inhibited IFNCSP binding to HepG2.2.15 cells and liver slices.
  
  
  CONCLUSIONS
  These results indicate that the molecular mechanisms responsible for IFN-CSP targeting involve binding to HSPG of hepatocytes and liver.
authors: luxuemei605@163.com.
authors: Xuemei
authors: X
authors: Lu
authors: Jie
authors: J
authors: Wang
authors: Xiaobao
authors: X
authors: Jin
authors: Yanting
authors: Y
authors: Huang
authors: Wenting
authors: W
authors: Zeng
authors: School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou 510006, People's Republic of China. zhujiayong1020@163.com.
authors: Jiayong
authors: J
authors: Zhu
mesh_terms: D000818:Animals; D001665:Binding Sites; D056945:Hep G2 Cells; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000008:administration & dosage* / Q000737:chemistry; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010961:Plasmodium / Q000737:chemistry; D015800:Protozoan Proteins / Q000008:administration & dosage* / Q000737:chemistry; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry
chemical_list: D019812:Heparan Sulfate Proteoglycans; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D015800:Protozoan Proteins; D011994:Recombinant Proteins