Functional selection of hepatitis C virus envelope E2-binding Peptide ligands by using ribosome display.
Small peptides that inhibit the hepatitis C virus (HCV) at the stage of viral entry have the potential to serve as attractive antiviral drugs. Ribosome display is a cell-free system for in vitro selection of peptides from large random peptide libraries. Thus, we utilized a ribosome display library technique for affinity selection of HCV envelope protein E2-binding peptide ligands. Through 13 rounds of selection, the ribosome display system generated high-affinity 12-mer peptides, and the selected peptide PE2D (MARHRNWPLVMV) demonstrated the highest specificity and affinity to the HCV E2 protein. Furthermore, amino acids 489 to 508 (YPPRPCGIVPAKSVCGPVYC) of E2 were identified as crucial for binding to PE2D. The selected peptides, especially PE2D, not only dramatically blocked E2 protein binding to hepatocytes but also dramatically inhibited HCV cell culture (HCVcc) entry into hepatocytes. HCVcc and HCV particles from HCV patient serum samples could also be specifically captured using PE2D. Our study demonstrates that the newly selected peptide ligand PE2D holds great promise for developing a new molecular probe, a therapeutic drug specifically for HCV, or an early-diagnostic reagent for HCV surface envelope antigen E2.
Chen
Fang
F
Department of Immunology, Wuhan University School of Medicine, Donghu Road 165, Wuhan 430071, People's Republic of China.
Zhao
Yinglan
Y
Liu
Min
M
Li
Dongqing
D
Wu
Hongyan
H
Chen
Haidan
H
Zhu
Yongzhe
Y
Luo
Fengling
F
Zhong
Jin
J
Zhou
Yidan
Y
Qi
Zhongtian
Z
Zhang
Xiao-Lian
XL
D000998:Antiviral Agents / Q000378:metabolism* / Q000494:pharmacology; D002352:Carrier Proteins; D045744:Cell Line, Tumor; D016174:Hepacivirus / Q000187:drug effects / Q000378:metabolism*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D008024:Ligands; D015335:Molecular Probes / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D012270:Ribosomes / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism*
D000998:Antiviral Agents; D002352:Carrier Proteins; D008024:Ligands; D015335:Molecular Probes; D019151:Peptide Library; D010455:Peptides; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus