Characterization of plasma membrane-associated proteins from Aedes albopictus mosquito (C6/36) cells that mediate West Nile virus binding and infection.
This study isolated and characterized the West Nile virus (WNV) putative receptor molecule(s) from Aedes albopictus mosquito (C6/36) cells. The binding of WNV to C6/36 cells was saturated with 5000 particles per cell. The entry of WNV into C6/36 cells was strongly inhibited when pretreated with proteinase K and to a lesser extent with sodium periodate. However, pretreatment of C6/36 cells with phospholipases, glycosidases, heparinases and neurimidase had no effect on virus entry. By using virus overlay protein blot assay, WNV was observed to bind to the 140-kDa, 95-kDa, 70-kDa and 55-kDa plasma membrane-associated molecules isolated from C6/36 cells. Murine antibodies generated against the 95-kDa and 70-kDa membrane proteins effectively blocked WNV, Japanese encephalitis virus (JEV) and Dengue virus (DV) serotype 2 infection in C6/36 cells. In addition, the binding of the recombinant-WNV envelope domain III protein to C6/36 cells can be inhibited by the anti-95-kDa and anti-70-kDa membrane protein antibodies. These data strongly supported the possibility that the 95-kDa and 70-kDa plasma membrane-associated proteins are part of a receptor complex for mosquito-borne flaviviruses (WNV, JEV and DV) on mosquito cells.
Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, 117597, Singapore.
J J H
JJ
Chu
P W H
PW
Leong
M L
ML
Ng
D000330:Aedes / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D009033:Culicidae; D015870:Gene Expression; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D014901:West Nile Fever / Q000378:metabolism*; D014902:West Nile virus / Q000502:physiology*
D008565:Membrane Proteins