Virus binding and CDHR3 glycosylation.

Lysates from HeLa cells transfected (A) or transduced (B) with full-length cDNAs encoding Y529 or C529 CDHR3 proteins were reacted with C15 virus in the presence or absence of PNGaseF or neuraminidase as described in Methods. Immunoprecipitation and protein detection used an α-CDHR3 mAb reactive with the cytoplasmic domain. Captured virus was quantitated as in Fig 1. (C) As in B, transduced cell lysates were treated (or not) with biotinylated Sambucus nigra (S) or Maackia amurensis (M) lectins before extraction with streptavidin agarose beads and protein detection with α-CDHR3. (D) Plated stably transduced HeLa cells expressing FLAG-Y529 or C529 CDHR3 were biotinylated and labeled cell surface proteins were isolated with streptavidin agarose beads.