Heat shock protein 70 on Neuro2a cells is a putative receptor for Japanese encephalitis virus.
Japanese encephalitis virus (JEV) envelope (E) protein has been shown to play a critical role in attachment to cells. However, the receptor interacting with envelope protein has not been conclusively identified. Using mouse neuroblastoma (Neuro2a) cells and purified JEV-E protein in 'Virus Overlay Protein Binding Assay' followed by MALDI-TOF analysis, we identified 'heat shock protein 70' (Hsp70) as a possible receptor for JEV. Indirect immunofluorescence and flow-cytometry analysis demonstrated localization of Hsp70 on Neuro2a cell surface. Co-immunoprecipitation followed by Western blot analysis reconfirmed the interaction between Hsp70 and JEV-E protein. Further, anti-Hsp70 polyclonal-antibodies were able to block JEV entry into Neuro2a cells. Additionally, using the bioinformatic tool - FTDOCK, docking between the proteins was performed. Amongst six interacting structural poses studied one pose involving RGD motif on JEV-E and leucine(539) on Hsp70 displayed stable interaction. These observations indicate that Hsp70 serves as putative receptor for JEV in Neuro2A cells.
Das
Soma
S
Department of Neurovirology, National Institute of Mental Health and Neurosciences, Bangalore, India.
Laxminarayana
Suhas Venkataramana
SV
Chandra
Nagasuma
N
Ravi
Vasanthapuram
V
Desai
Anita
A
D000818:Animals; D016367:Baculoviridae / Q000254:growth & development / Q000378:metabolism; D045744:Cell Line, Tumor; D004664:Encephalitis Virus, Japanese / Q000276:immunology / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D009474:Neurons; D011485:Protein Binding; D011991:Receptors, Virus / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D053585:Virus Attachment; D053586:Virus Internalization
D018840:HSP70 Heat-Shock Proteins; D011991:Receptors, Virus; D014764:Viral Proteins