Identification of a heparin binding peptide from the Japanese encephalitis virus envelope protein

The flavivirus envelope protein is the dominant antigen in eliciting neutralizing antibodies and plays an important role in inducing immunologic responses in the infected host. It has been shown that highly sulfated forms of heparin sulfate can bind to the envelope protein and are involved in flavivirus infection. Among the three structural domains, domain III is the major antigenic domain of the envelope protein. We have prepared an extended form of the JEV domain III protein with residues ranging from 261 to 402 and determined its heparin binding sites. Based on NMR, fluorescence spectroscopy, and site-directed mutagenesis studies, we have identified that only the N-terminal region (residues 279-293) and some spatially adjacent residues of JEV domain III are involved in heparin binding. Moreover, a synthetic peptide corresponding to this region also demonstrates strong affinity to heparin. Our results provide a basis for further understanding the interactions of flaviviruses and glycosaminoglycans on the host cell surfaces.