A chemical lipid modification of recombinant preS antigen to study the mechanism of HBV attachment to the host cell.
Surface antigen preS of Hepatitis B virus plays fundamental roles in mediating receptor recognition and virus internalization. Myristoylation at N-terminal Gly(2) residue of preS is essential for viral attachment and infectivity. A number of myristoylated proteins have been shown to undergo a conformational change (myristoyl switch) that alters their affinity to cell membrane. However, there is little knowledge about what effect this fatty acylation contributes in virus-host cell interaction. Here we demonstrated a new method for lipid modification of recombinant preS protein at N-terminal residue 2 with alkylating chemicals. Modified preS was able to inhibit HBV penetrating into HepG2 cells with an increased efficiency compared to unmodified form. Flow cytometric analysis indicated that lipid modification enhanced the binding affinity of preS to hepatocytes, but not resulting from hydrophobic interaction. CD analysis further revealed a conformational change of modified preS in the presence of membrane mimetics. These findings imply that the conformation transition induced by fatty acylation is important for efficient attachment of virus to cell receptors, and this method of chemical lipid modification provides a basis for designing therapeutic inhibitors to Hepatitis B virus.
Chen
Bin
B
National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
Lian
Min
M
Xu
Shili
S
Luo
Ming
M
Zheng
Xiaofeng
X
D000215:Acylation; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D005434:Flow Cytometry; D006514:Hepatitis B Surface Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006515:Hepatitis B virus / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology; D006801:Humans; D008055:Lipids / Q000494:pharmacology; D019814:Myristic Acid / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053585:Virus Attachment / Q000187:drug effects
D006514:Hepatitis B Surface Antigens; D008055:Lipids; D011498:Protein Precursors; D011994:Recombinant Proteins; D019814:Myristic Acid