从植物种子中纯化和鉴定两种电压依赖性阴离子通道亚型
Purification and characterization of two voltage-dependent anion channel isoforms from plant seeds.
作者信息
Abrecht H, Wattiez R, Ruysschaert J M, Homblé F
机构信息
Laboratoire de Physiologie Végétale, Faculté des Sciences, Université Libre de Bruxelles, B-1050 Brussels, Belgium.
出版信息
Plant Physiol. 2000 Nov;124(3):1181-90. doi: 10.1104/pp.124.3.1181.
Abstract
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.
摘要
从小扁豆（兵豆）和菜豆吸胀的种子中分离出线粒体。我们使用羟基磷灰石，在单个纯化步骤中从去污剂溶解的线粒体中共同纯化了两种电压依赖性阴离子通道。菜豆的两种同工型通过在4M尿素中的层析聚焦色谱法分离，且通道活性没有任何损失。将每种同工型重组到二植酰磷脂酰胆碱平面脂质双分子层中后，对其通道活性进行了表征。两种同工型均形成大电导通道，这些通道对阴离子有轻微选择性，并显示出阳离子选择性亚态。
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