Plant Physiol, November 2000, Vol. 124, pp. 1181-1190
Purification and Characterization of Two Voltage-Dependent Anion
Channel Isoforms from Plant Seeds1
Helge
Abrecht,
Ruddy
Wattiez,
Jean-Marie
Ruysschaert, and
Fabrice
Homblé*
Laboratoire de Physiologie Végétale (H.A.,
F.H.) and Laboratoire de Chimie Physique des Macromolécules aux
Interfaces (J.-M.R.), Faculté des Sciences, Université
Libre de Bruxelles, B-1050 Brussels, Belgium; and Laboratoire de
Biochimie, Faculté des Sciences, Université de
Mons-Hainaut, B-7000 Mons, Belgium (R.W.)
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.
1
This work was supported by a European Commission
Research Training grant (no. ERBFMBICT971977 to H.A.). F.H. is a
Research Director from the National Fund for Scientific Research (Belgium).
*
Corresponding author; e-mail fhomble{at}ulb.ac.be; fax
32-2- 650-5382.
© 2000 American Society of Plant Physiologists
