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. 2004 Sep;78(18):9666–9674. doi: 10.1128/JVI.78.18.9666-9674.2004

FIG. 6.

FIG. 6.

The presence of the measles virus H (MVH) protein in the ER slows or prevents the complex glycosylation of SLAM but not CD46. Human 293Tad embryonic kidney cells were transfected with pcDNA3.1-SLAM-HA3 and either pCG, pCG-H, or pCG-HER. At 36 h posttransfection, cells were lysed and immunoprecipitated with anti-H, anti-CD46, or anti-HA tag antibodies. The resulting samples were left untreated (−) or treated with Endo H or PNGaseF (+). The samples were subjected to SDS-PAGE; transferred to nitrocellulose; probed with an anti-HA antibody, anti-H antibody, or anti-CD46 antibody; and detected with a peroxidase-conjugated secondary antibody by enhanced chemiluminescence. (A) Lysates were immunoprecipitated (IP) with an anti-HA antibody, and the blots were probed with an anti-HA tag antibody. Expression of H or HER inhibits complex glycosylation of SLAM and maintains its sensitivity to Endo H. (B) Lysates were immunoprecipitated with a mouse monoclonal anti-CD46 antibody that recognized endogenous CD46. The blots were probed with a rabbit polyclonal anti-CD46 antibody. Expression of H or HER did not inhibit the complex glycosylation of CD46, and the glycoprotein exhibited little or no sensitivity to Endo H. (C) Lysates were immunoprecipitated with a mouse monoclonal anti-MVH antibody, and the blots were probed with an anti-HA antibody. The SLAM receptor coprecipitates with MVH proteins, and HER expression prevents the complex glycosylation of SLAM and maintains its sensitivity to Endo H. (D) Lysates were immunoprecipitated with an anti-HA antibody, and the blots were probed with a rabbit polyclonal anti-MVH antibody. MVH coprecipitates with SLAM and is partially sensitive to Endo H. (E) 293Tad cells were transfected with only pCG, pCG-H, or pCG-HER. Lysates were immunoprecipitated with a mouse monoclonal anti-MVH antibody, and the blots were probed with a rabbit polyclonal anti-CD46 antibody as in panel B. Endogenous CD46 does not coprecipitate with MVH. (F) 293Tad cells were transfected with only pCG, pCG-H, or pCG-HER. Lysates were immunoprecipitated with a mouse monoclonal anti-CD46 antibody, and the blots were probed with a rabbit polyclonal anti-MVH antibody as in panel D. MVH does not coprecipitate with endogenous CD46.