The mitochondrial voltage-dependent channel, VDAC, is modified asymmetrically by succinic anhydride
In the accompanying paper, succinic anhydride was shown to react with the outer mitochondrial membrane channel-forming protein, VDAC, resulting in the loss of its voltage dependence. In this paper, the anhydride was added to VDAC held in a particular conformational state by means of an applied electric field. VDAC was inserted into the membranes from thecis side and the anhydride was added either to thecis ortrans side. Channels modified in the open state behaved similarly whether anhydride was added to thecis ortrans side. Modifications of VDAC in either of the two closed states did not. Modifications resulting in the loss of voltage-dependence occurred primarily when anhydride was added to the negative side of the membrane irrespective of which closed state the VDAC was in indicating that the accessibility of the gating charges alternated between thecis andtrans sides as the channel's conformation was changed from one closed state to the other. Despite the pronounced asymmetry, in general the resulting channels behaved in the same way in response to either positive or negative fields. A model consistent with the results is presented which proposes that the same gating charges are responsible for channel closure at both positive and negative fields.
Key Wordsvoltage-gating protein modification mechanism planar lipid membrane outer membrane ion transport
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