CHARACTERIZATION OF THE MAJOR INTEGRAL PROTEIN OF VACUOLAR MEMBRANE
CHARACTERIZATION OF THE MAJOR INTEGRAL PROTEIN OF VACUOLAR MEMBRANE
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DOI：
10.1104/pp.98.4.1248
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发表时间：
1992-04-01
期刊：
影响因子：
7.4
通讯作者：
MAESHIMA, M
中科院分区：
文献类型：
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作者：
MAESHIMA, M
萝卜（Raphanus sativus）主根液泡膜中含有大量的蛋白质，约为23千道尔顿，占总膜蛋白的25%以上。该蛋白被纯化并进行了表征，暂命名为VM 23。即使在1%十二烷基硫酸钠存在的情况下，vm23在高温下也容易聚集。在0.1% Triton X-100的存在下，聚丙烯酰胺凝胶电泳估计vm23的表观分子大小约为400千道尔顿。用氯仿甲醇从液泡膜中部分提取vm23，表明其具有较高的疏水性。疏水性羧基改性剂N，N'-双环己基碳二胺共价结合到vm23上。结果表明，vm23可能是一个与质子输运耦合的二次输运系统。抗萝卜vm23抗体能与绿豆（Vigna radiata）、蓖麻（Ricinus communis）下胚轴和南瓜（Cucurbita moschata）上胚轴空泡膜上的主要蛋白发生反应，而与甜菜（Beta vulgaris）主根空泡膜上的主要蛋白不发生反应。膜超声处理后，在蔗糖密度梯度离心的条件下，VM 23与空泡H+-焦磷酸酶结合，表明其与空泡膜相关。
The vacuolar membrane of radish (Raphanus sativus) taproot contained a large quantity of a protein of 23 kilodaltons that accounted for more than 25% of the total membrane proteins. The protein, tentatively named VM 23, was purified and characterized. VM 23 tends to aggregate at high temperature even in the presence of 1% sodium dodecyl sulfate. The apparent molecular size of VM 23 was estimated to be about 400 kilodaltons by polyacrylamide gel electrophoresis in the presence of 0.1% Triton X-100. VM 23 was partially extracted from the vacuolar membranes with chloroform:methanol, indicating its high hydrophobicity. The hydrophobic carboxyl modifier N,N'-dicyclohexylcarbodimide bound covalently to VM 23. The results suggest that VM 23 may act as a secondary transport system coupled with the proton transport. The antibody against radish VM 23 reacted with the major proteins in the vacuolar membranes of mung bean (Vigna radiata) and castor bean (Ricinus communis) hypocotyls and pumpkin (Cucurbita moschata) epicotyl, but not with that of sugar beet (Beta vulgaris) taproot. VM 23 comigrated with vacuolar H+-pyrophosphatase on sucrose density gradient centrifugation after sonication of membranes, indicating that it is associated with the vacuolar membrane.