Salt D E, Wagner G J
Agronomy Department, University of Kentucky, Lexington 40546-0091.
J Biol Chem. 1993 Jun 15;268(17):12297-302.
Cadmium accumulates in the vacuole of plant cells, but the mechanism driving its transport across the vacuole membrane is not understood. Here we present evidence for Cd2+ transport via a Cd2+/H+ antiport activity into tonoplast-enriched vesicles isolated from oat roots. Experimentally, accumulation of Cd2+ into vesicles could be driven by delta pH generated by either V-type ATPase or artificially using nigericin to exchange K+ and H+ in K(+)-loaded vesicles. When tonoplast-enriched vesicles were separated on a linear sucrose gradient, NO3(-)-sensitive ATPase, total MgATPase, and delta pH-dependent Cd2+ transport equilibrated at 1.11 g/ml, the density of tonoplast membrane. Cd2+ accumulation in vesicles was accompanied by efflux of protons in a Cd2+ concentration-dependent manner characteristic of an antiport activity. The delta pH-dependent Cd2+ accumulation process showed saturation kinetics with a Km(app) of 5.5 microM. Thus the process is a candidate for transport of Cd2+ from the cytoplasm to the vacuolar sap under conditions of low as well as high Cd2+ exposure.
镉在植物细胞的液泡中积累,但其跨液泡膜运输的机制尚不清楚。在此,我们提供证据表明,镉离子通过镉离子/氢离子反向转运活性进入从燕麦根部分离出的富含液泡膜的囊泡。实验表明,囊泡中镉离子的积累可由V型ATP酶产生的ΔpH驱动,或通过在装载钾离子的囊泡中使用尼日利亚菌素人工交换钾离子和氢离子来驱动。当富含液泡膜的囊泡在线性蔗糖梯度上分离时,对硝酸根敏感的ATP酶、总镁ATP酶以及依赖于ΔpH的镉离子转运在1.11克/毫升(液泡膜的密度)处达到平衡。囊泡中镉离子的积累伴随着质子以反向转运活性特有的镉离子浓度依赖性方式流出。依赖于ΔpH的镉离子积累过程呈现饱和动力学,表观米氏常数(Km(app))为5.5微摩尔。因此,在低镉和高镉暴露条件下,该过程是镉离子从细胞质运输到液泡汁液的一个候选机制。