Purification of a HeLa cell receptor protein for group B coxsackieviruses.
Coxsackievirus B3 (CB3) firmly attaches to HeLa cells, forming a specific complex between the virus and its receptor on the cell surface. We extracted this virus-receptor complex (VRC) with the detergents sodium deoxycholate and Triton X-100. The VRC was identified by its sedimentation coefficient (140S), which was less than that of virions (155S). Formation of the VRC from cell lysates and 3H-CB3 occurred with the same specificity as did attachment of virions to cells, in that its formation was blocked by unlabeled CB3 but not by poliovirus. The VRC was purified 30,000-fold by differential and sucrose gradient centrifugation. Iodination with Na125I revealed that the purified VRC consisted of the normal CB3 proteins and one additional protein (RP-a) with an approximate molecular weight of 49,500. RP-a was eluted from the VRC and was shown to rebind with CB3 and CB1 virions but not with poliovirus type 1. We propose that Rp-a is a protein in the plasma membrane receptor complex which is responsible for the specific recognition and binding of the group B coxsackieviruses.
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D002499:Centrifugation, Density Gradient; D029822:Enterovirus B, Human / Q000502:physiology*; D006367:HeLa Cells / Q000032:analysis*; D006801:Humans; D007455:Iodine / Q000378:metabolism; D011991:Receptors, Virus / Q000302:isolation & purification*; D014764:Viral Proteins / Q000378:metabolism
D011991:Receptors, Virus; D014764:Viral Proteins; D007455:Iodine