Novel cell adhesive glycosaminoglycan-binding proteins of Japanese encephalitis virus.
Glycosaminoglycans (GAGs) are present in the extracellular matrix and/or tissue cell surface and, by binding to specified GAG-binding proteins, control many important cellular functions. Some animal viruses had evolved to use GAGs as part of their strategy to invade host cells. In this study, two putative GAG-binding proteins were identified from the E protein sequence of the live-attenuated strain CH2195LA of Japanese encephalitis virus (JEV): (i) the first GAG-binding region at residues from E-279 to E-297 (279KLTSGHLKCRLKMDKLALK297) and (ii) the second GAG-binding region at residues from E-397 to E-416 (397KAGSTLGKAFFSTTLKGAQR416). Four recombinant proteins with or without these two GAG-binding regions were expressed in Escherichia coli and purified to examine their GAG-binding properties. The first GAG binding region was demonstrated to exhibit a higher affinity in heparin-Sepharase column. Dose-dependent increases of BHK-21 cell binding were also demonstrated by cell binding enzyme-linked immunosorbent assay (ELISA). Immobilized on glass coverslips, the GAG-binding recombinant protein of JEV promoted BHK-21 cell adhesion and proliferation. The present studies demonstrate the recombinant GAG-binding proteins of JEV stimulate cell adhesive and proliferation with a potential for applications in tissue engineering.
Wu
Suh-Chin
SC
Institute of Biotechnology, Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan, Republic of China. scwu@life.nthu.edu.tw
Chiang
Jeng-Ru
JR
Lin
Cheng-Wen
CW
D000595:Amino Acid Sequence; D000818:Animals; D001672:Biocompatible Materials; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D023822:Tissue Engineering; D014764:Viral Proteins / Q000737:chemistry*
D001672:Biocompatible Materials; D006025:Glycosaminoglycans; D011994:Recombinant Proteins; D014764:Viral Proteins; D006493:Heparin; D012685:Sepharose