Affixing N-terminal ?-helix to the wall of the voltage-dependent anion channel does not prevent its voltage gating.

ABSTRACT: The voltage-dependent anion channel (VDAC) governs the free exchange of ions and metabolites between the mitochondria and the rest of the cell. The three-dimensional structure of VDAC1 reveals a channel formed by 19 ?-strands and an N-terminal ?-helix located near the midpoint of the pore. The position of this ?-helix causes a narrowing of the cavity, but ample space for metabolite passage remains. The participation of the N-terminus of VDAC1 in the voltage-gating process has been well established, but the molecular mechanism continues to be debated; however, the majority of models entail large conformational changes of this N-terminal segment. Here we report that the pore-lining N-terminal ?-helix does not undergo independent structural rearrangements during channel gating. We engineered a double Cys mutant in murine VDAC1 that cross-links the ?-helix to the wall of the ?-barrel pore and reconstituted the modified protein into planar lipid bilayers. The modified murine VDAC1 exhibited typical voltage gating. These results suggest that the N-terminal ?-helix is located inside the pore of VDAC in the open state and remains associated with ?-strand 11 of the pore wall during voltage gating.

SUBMITTER: Teijido O 

PROVIDER: S-EPMC3322836 | biostudies-other | 2012 Mar

REPOSITORIES: biostudies-other