Purification and characterization of two voltage-dependent anion channel isoforms from plant seeds

Mitochondria were isolated from imbibed seeds of lentil (Lens culinaris) an d Phaseolus vulgaris. We copurified two voltage-dependent anion channel fro m detergent solubilized mitochondria in a single purification step using hy droxyapatite. The two isoforms from P. vulgaris were separated by chromatof ocusing chromatography in 4 M urea without any loss of channel activity, Ch annel activity of each isoform was characterized upon reconstitution into d iphytanoyl phosphatidylcholine planar lipid bilayers. Both isoforms form la rge conductance channels that are slightly anion selective and display cati on selective substates.