Booth J W, Guidotti G
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
J Biol Chem. 1997 Aug 15;272(33):20408-13. doi: 10.1074/jbc.272.33.20408.
The vacuole of the yeast Saccharomyces cerevisiae is a major storage compartment for phosphate. We have measured phosphate transport across the vacuolar membrane. Isolated intact vacuoles take up large amounts of added [32P]phosphate by counterflow exchange with phosphate present in the vacuoles at the time of their isolation. The bidirectional phosphate transporter has an intrinsic dissociation constant for phosphate of 0.4 mM. Exchange mediated by this carrier is faster than unidirectional efflux of phosphate from the vacuoles. The transporter is highly selective for phosphate; of other anions tested, only arsenate is also a substrate. Transport is strongly pH-dependent with increasing activity at lower pH. Similar phosphate transport behavior was observed in right-side-out vacuolar membrane vesicles.
酿酒酵母的液泡是磷酸盐的主要储存区室。我们测量了磷酸盐跨液泡膜的转运。分离得到的完整液泡通过与分离时液泡中存在的磷酸盐进行逆流交换,摄取大量添加的[32P]磷酸盐。双向磷酸盐转运体对磷酸盐的内在解离常数为0.4 mM。由该载体介导的交换比磷酸盐从液泡中的单向流出更快。该转运体对磷酸盐具有高度选择性;在所测试的其他阴离子中,只有砷酸盐也是一种底物。转运强烈依赖于pH值,在较低pH值下活性增加。在外翻的液泡膜囊泡中也观察到类似的磷酸盐转运行为。