Fig. 6. Physical association of Ccz1p with Ypt7p. Total extracts from yeast cells were immunoprecipitated with anti-HA antibody as described in Materials and Methods. Total extracts, 3 µl (A), and immunoprecipitates from 1 ml of extracts (B) were analyzed by SDS-PAGE and immunoblotting with anti-HA antibody (upper panel) to detect Ccz1-HAp and with antiserum to Ypt7p (lower panel). The band corresponding for Ccz1-Hap is not detectable even for 20 µl of extracts loaded on gel. Lanes represent preparations from strains: (1) ypt7; (2) ccz1 [YPT7]; (3) ccz1 [YPT7/CCZ1-HA]; (4) ccz1 [ypt7^K127E /CCZ1-HA]; (5) ccz1 [ypt7^Q68L/CCZ1-HA]; (6) ccz1 [ypt7^T22N /CCZ1-HA]. The double bands visible for Ypt7^K127Ep probably represent geranylgeranylated and unmodified forms.