Figure 5

Effects of the KK, TK, and KKTK substitutions on the association of Boi-PH with phospholipids. A). Shown are immunoblots, probed with anti-Boi1 antibodies, of aliquots from yeast cytosolic extracts containing either wild-type or mutant versions of Boi1-PH, after incubation with either no vesicles (lanes 1, 4, 7, and 10), vesicles containing only PS (lanes 2, 5, 8, and 11), or vesicles composed of a 1:20 (mass:mass) mixture of PIP₂ and PS (lanes 3, 6, 9, and 12). B). The four panels show immunoblots, probed with anti-Boi1 antibodies, of supernatant (S) and pellet (P) fractions, resulting after centrifugation, of yeast cytosolic extracts containing either wild-type Boi1-PH (top panel), Boi1-PH(KK) (second panel), Boi1-PH(TK) (third panel), or Boi1-PH(KKTK) (bottom panel), with either no vesicles (lanes 1 and 2), vesicles composed only of PS (lanes 3 and 4), or vesicles composed of a 1:20 (mass:mass) mixture of PIP₂ and PS (lanes 5 and 6).