FIGURE 8.
N-Glycosylation is required for the trafficking of PRiMA-linked mixed chicken-human AChE tetramer. A, HEK293T cells were transfected with the cDNAs encoding AChETWT or AChETN296Q/N381Q/N495Q together with cAChET and PRiMA. After 48 h the transfected cells were subjected to subcellular fractionation, and the distribution of ER and Golgi/PM subcellular compartments was determined by immunoblotting as in Fig. 6A. The distribution of AChET and cAChET proteins was analyzed by immunoblotting with the anti-AChE antibodies. B, protein extracts from the total lysate and ER-enriched and Golgi/PM-enriched fractions of the transfected cells from A were immunoprecipitated (IP) by either anti-human AChE antibody or anti-chicken AChE antibody (Ab). The immunoprecipitated complexes were analyzed by SDS-PAGE and Western blotting with anti-human AChE and anti-chicken AChE antibodies as indicated. Representative gels from three independent experiments are shown.