Reuveni M, Bennett A B, Bressan R A, Hasegawa P M
Center for Plant Environmental Stress Physiology-Department of Horticulture and Department of Entomology, Purdue University, West Lafayette, Indiana 47907.
Plant Physiol. 1990 Oct;94(2):524-30. doi: 10.1104/pp.94.2.524.
Tonoplast enriched membrane vesicle fractions were isolated from unadapted and NaCl (428 millimolar) adapted tobacco cells (Nicotiana tabacum L. var Wisconsin 38). Polypeptides from the tonoplast enriched vesicle fractions were separated by SDS-PAGE and analyzed by Western blots using polyclonal antibodies to the 70 kilodalton subunit of the red beet tonoplast H(+)-ATPase. These antibodies cross-reacted exclusively to a tobacco polypeptide of an apparent molecular weight of 69 kilodaltons. The antibodies inhibited ATP-dependent, NO(3) (-) sensitive H(+) transport into vesicles in tonoplast enriched membrane fractions from both unadapted and NaCl adapted cells. The relative H(+) transport capacity per unit of 69 kilodalton subunit of the tonoplast ATPase of vesicles from NaCl adapted cells was fourfold greater than that observed for vesicles from unadapted cells. The increase in specific H(+) transport capacity after adaptation was also observed for ATP hydrolysis.
从未适应和经428毫摩尔氯化钠适应的烟草细胞（烟草品种Wisconsin 38）中分离出富含液泡膜的膜泡组分。通过SDS-PAGE分离富含液泡膜的膜泡组分中的多肽，并用针对红甜菜液泡膜H(+)-ATPase 70千道尔顿亚基的多克隆抗体进行Western印迹分析。这些抗体仅与一种表观分子量为69千道尔顿的烟草多肽发生交叉反应。这些抗体抑制了未适应和经氯化钠适应的细胞中富含液泡膜的膜组分中囊泡的ATP依赖性、NO(3) (-) 敏感的H(+) 转运。与未适应细胞的囊泡相比，经氯化钠适应的细胞的囊泡中每单位69千道尔顿液泡膜ATPase亚基的相对H(+) 转运能力大四倍。适应后特异性H(+) 转运能力的增加在ATP水解中也有观察到。
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