FIG. 5.

A mutant with a serine-to-threonine change at residue 507 in HeV-G gains ephrinB3 binding and viral entry abilities comparable to those of NiV. (A) sNiV-G and sHeV-G with residue T507 (sH-507T) or S507 (sH-507S), used at 2 nM, 0.4 nM, and 0.1 nM concentrations, were allowed to bind independently to CHO-B2 or CHO-B3 cells. The amount of binding was assessed by flow cytometry using anti-HA monoclonal antibodies and R-phycoerythrin-conjugated anti-mouse IgG. Data are the averages of triplicates ± SD. (B) Henipavirus envelope glycoproteins, namely, NiV-F/G (NiVpp) or HeV-F/G with residue 507T or 507S (H-507T or H-507S, respectively), were pseudotyped onto VSV reporter viruses. NiVpp, H-507T, and H-507S virus particles were run in a reducing SDS-PAGE gel and detected by anti-HA (αHA) Western blotting to detect G. Note that HeV-G always runs slightly higher on the gel than NiV-G. (C) Fivefold serial dilutions were made with NiVpp, H-507T, and H-507S VSV reporter viruses and subsequently used to infect CHO-B2 and CHO-B3 cells at similar concentrations, based on the amount of G incorporated onto the viral particles (B). Luc reporter activity was measured in RLU. Data are the averages for three independent experiments ± SD.