Isolation and characterization of the mitochondrial channel, VDAC, from the insect Heliothis virescens

A 31 kDa voltage-dependent anion-selective channel (VDAC) protein was purified from the insect Heliothis virescens (tobacco budworm, denoted TBW) using an alkali extraction and filtration procedure and was characterized by SDS-PAGE, amino acid sequencing, biophysical properties and immunocytochemistry. The N-terminal sequence has highest identity with VDACs from mammals (50-66%) followed by plants (34-41%) and lower eukaryotes (30-34%). Reconstitution in planar phospholipid membranes yielded properties typical of VDACs from other organisms including a single-channel conductance of 4.1 nS (in 1 M KCl), closure in response to positive and negative transmembrane voltage, and a reversal potential of 11.8 mV indicating anion selectivity in the open state. A polyclonal antiserum (R19) raised against gel-purified 31 kDa protein specifically labelled mitochondria and mitochondrial outer membranes in TBW flight muscle by light and electron microscope immunocytochemistry.