Fig. 2.

VopQ inhibits trans-SNARE complex formation. (A) At 0-, 10-, 20-, and 45-min intervals, ALP vacuole fusion reaction was incubated on ice, or one of the following fusion inhibitors was added: α-Sec17p, α-Ypt7p, α-Vam3p, or 200 nM VopQ. After 90 min, fusion was measured and compared with the fusion of an uninhibited fusion reaction. Error bars indicate SD from the mean; n = 3. (B) The quantitative docking assay was performed in the presence of 500 nM MBP, 2.8 µM Gdi1p and 11.4 µM Gyp1-46, or 500 nM MBP-VopQ. At least 130 clusters from 10 random fields were scored for each condition. (C) Trans-SNARE assays were performed without or with the following inhibitors: ice incubation, 1 µM Gdi1p/1 µM Gyp1-46, or 200 nM VopQ. (D) ALP vacuole fusion reactions were performed in the absence or presence of 200 nM MBP-VopQ. Recombinant 50 nM Vam7p SNARE, 20 nM HOPS complex, or α-Vam3p was added where indicated.