Department of Botany and Plant Sciences, University of California, Riverside, California 92521.
Plant Physiol. 1980 Oct;66(4):684-7. doi: 10.1104/pp.66.4.684.
Vacuoles were isolated from protoplasts of Nicotiana glutinosa by the method of Mettler and Leonard (Plant Physiol 1979 64: 1114-1120) with minor modifications so that the number of intact protoplasts contaminating the vacuole preparation was reduced to less than 1% (by number). Isopycnic centrifugation of a [(3)H]choline-labeled, sonicated vacuole preparation on linear 5 to 40% sucrose gradients indicated that tonoplast vesicles equilibrated at a density of about 1.12 grams per cubic centimeter. When tonoplast vesicles were isolated on discontinuous sucrose density gradients substrate specific ATPase activity was not found to be associated with this membrane fraction. These results are discussed in terms of the energetics of ion transport through the tonoplast membrane.
通过 Mettler 和 Leonard（植物生理学 1979 年 64 卷：1114-1120）的方法对烟草原生质体进行轻微修改后，从小麦原生质体中分离出液泡，使污染液泡制备物的完整原生质体数量减少到小于 1%（按数量计）。对用 [(3)H]胆碱标记并超声处理的液泡制剂进行等密度离心，在 5 至 40%蔗糖梯度上，表明质膜小泡在约 1.12 克/立方厘米的密度处达到平衡。当在不连续蔗糖密度梯度上分离质膜小泡时，未发现底物特异性 ATP 酶活性与该膜部分相关。这些结果是根据通过质膜的离子运输的能量学来讨论的。
