Purification and characterization of two voltage-dependent anion channel isoforms from plant seeds.
Mitochondria were isolated from imbibed seeds of lentil (Lens culinaris) and Phaseolus vulgaris. We copurified two voltage-dependent anion channel from detergent solubilized mitochondria in a single purification step using hydroxyapatite. The two isoforms from P. vulgaris were separated by chromatofocusing chromatography in 4 M urea without any loss of channel activity. Channel activity of each isoform was characterized upon reconstitution into diphytanoyl phosphatidylcholine planar lipid bilayers. Both isoforms form large conductance channels that are slightly anion selective and display cation selective substates.
Abrecht
H
H
Laboratoire de Physiologie Végétale, Faculté des Sciences, Université Libre de Bruxelles, B-1050 Brussels, Belgium.
Wattiez
R
R
Ruysschaert
J M
JM
Homblé
F
F
D000595:Amino Acid Sequence; D007887:Fabaceae / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D007473:Ion Channels / Q000737:chemistry / Q000302:isolation & purification*; D008051:Lipid Bilayers / Q000737:chemistry; D008928:Mitochondria / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010713:Phosphatidylcholines / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification; D010946:Plants, Medicinal; D018272:Porins / Q000737:chemistry / Q000302:isolation & purification*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D050994:Voltage-Dependent Anion Channels
D007473:Ion Channels; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010940:Plant Proteins; D018272:Porins; D020033:Protein Isoforms; D050994:Voltage-Dependent Anion Channels; C021016:1,2-diphytanoylphosphatidylcholine