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首页> 外文期刊>Cellular Physiology and Biochemistry >Role of vacuolar ATPase in the trafficking of renal type IIa sodium-phosphate cotransporter
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Role of vacuolar ATPase in the trafficking of renal type IIa sodium-phosphate cotransporter

机译：液泡ATPase在肾脏IIa型磷酸钠共转运蛋白运输中的作用

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摘要

Total body phosphate homeostasis is regulated by expression of type IIa sodium phosphate cotransporter (NpT2a) in the apical membrane (BBM) of renal proximal tubule cells. NpT2a expression is regulated by dietary phosphate and PTH but the mechanisms for trafficking of the protein are unknown. Based on 2D gel electrophoresis and mass spectroscopy data that changes in dietary phosphate stimulated changes in BBM expression of vacuolar H ~+-ATPase, we hypothesized that vacuolar H~+-ATPase plays a significant role in regulation of NpT2a in opossum kidney (OK) cells, a model for renal proximal tubule transport. Methods: Role of vacuolar H ~+-ATPase was studied in opossum kidney (OK) cells by examining the effect of inhibition of vacuolar H~+-ATPase on Pi uptake and NpT2a expression. Results: Pretreatment of OK cells with bafilomycin A1 and concanamycin A, inhibitors of vacuolar H~+-ATPases, blocked high phosphate- and PTH-induced degradation of NpT2a, but had no effect on high phosphate or PTH induced inhibition of sodium-dependent phosphate transport. Exposure of the cells to bafilomycin A_1 significantly decreased phosphate transport and apical membrane expression of NpT2a. Treatment with brefeldin A, an inhibitor of Golgi transport, decreased phosphate transport and apical membrane expression of NpT2a while treatment of cells with both brefeldin A and bafilomycin A_1 had no additive effect. Conclusion: We conclude that vacuolar H~+-ATPase plays a significant role in exocytosis of NpT2a into the apical membrane and in degradation of NpT2a but has no role in endocytosis.
机译：总体磷酸盐体内稳态受肾脏近端小管细胞顶膜（BBM）中IIa型磷酸钠共转运蛋白（NpT2a）表达的调节。 NpT2a的表达受饮食中的磷酸盐和PTH调节，但蛋白运输的机制尚不清楚。根据2D凝胶电泳和质谱数据（饮食中磷酸盐的变化刺激液泡H〜+ -ATPase的BBM表达变化），我们假设液泡H〜+ -ATPase在负鼠肾NpT2a的调节中起重要作用（确定）细胞，肾近端小管运输的模型。方法：通过观察液泡H〜+ -ATP酶抑制Pi摄取和NpT2a表达的作用，研究液泡H〜+ -ATPase在负鼠肾（OK）细胞中的作用。结果：巴氟霉素A1和伴刀豆球蛋白A抑制液泡H〜+ -ATPases的OK细胞预处理，阻断了高磷酸盐和PTH诱导的NpT2a降解，但对高磷酸盐或PTH诱导的钠依赖性磷酸盐抑制没有作用运输。细胞暴露于bafilomycin A_1显着降低了NpT2a的磷酸盐转运和顶膜表达。布雷菲德菌素A（一种高尔基体转运抑制剂）可以降低磷酸盐转运和NpT2a的顶膜表达，而布雷菲德菌素A和巴菲洛霉素A_1的细胞均无累加作用。结论：我们得出的结论是液泡H〜+ -ATPase在NpT2a胞吐进入顶膜和NpT2a降解中起重要作用，但在胞吞作用中不起作用。

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