QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 9. The PT pore increases diffusibility of intermembrane space proteins but is not involved in the action of tBid at low concentrations. CyP-D(+) mitochondria were used throughout. (A) Calcein-loaded mitochondria were incubated in SRM containing either 400 µM Ca²⁺ (400 Ca) with/without 1 µM CSA, or 1 µM buffered free Ca²⁺ (1 Ca) with/without 20 nM tBid. The Ca²⁺ buffer comprised 5 mM EGTA and 4 mM CaCl₂. After incubation for the time specified, mitochondria were sedimented and calcein in the supernatant was determined fluorometrically. (B) Calcein-loaded mitochondria were incubated in SRM containing 400 µM Ca²⁺, 1 µM free Ca²⁺ and 20 nM tBid, as indicated. Cytochrome c release was measured at the times specified. The data of A and B are representative of three such experiments. (C) The two-step procedure was used to determine the diffusibility rate constants for cytochrome c (white columns) and Smac (black columns) in the presence of 20 nM tBid. Other additions were 400 µM Ca²⁺, 1 µM buffered free Ca²⁺ and 1 µM CSA, as indicated. Means ± s.e.m. (four experiments).

Return to article