Comparison of the transport properties of three plant sucrose carriers expressed in Xenopus oocytes

Jing-Jiang Zhou, A. Miller
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引用次数: 10

Abstract

Previous work has shown different patterns of expression for the three plant sucrose transporters PmSUC1, PmSUC2 and AtSUC1. Each carrier may therefore have different properties appropriate for the transport function at each location in the plant. To test this hypothesis, a detailed characterisation of each carrier was performed by expressing the protein in Xenopus oocytes. All three carriers were functionally active in oocytes and transported both sucrose and maltose with the sugars eliciting inward currents, which increased at more negative membrane voltages. This effect was greater for AtSUC1 and PmSUC1 than for PmSUC2. The affinities of the carriers for each sugar were different, with PmSUC2 having a 10-fold higher Km for sucrose compared to AtSUC1 and PmSUC1. However, these values for all three carriers were dependent on the external pH and membrane voltage. For both PmSUC1 and PmSUC2 the maximal sugar transport was achieved at external pH values of 6.5-7.5, values which do not appear to be consistent with a proton-coupled mechanism of transport. However, the normal saline used for oocyte experiments contains relatively high concentrations of NaCl that could interfere with the function of the carrier protein. To test this idea the sucrose-elicited currents were measured in saline containing lower concentrations of these ions. Sodium, but not chloride, ions modified the sugar transport activities of the carriers and these effects were different for each carrier, inhibiting AtSUC1 and PmSUC1, but stimulating PmSUC2. The properties of each carrier are discussed in relation to their expression in oocytes and what is known about their pattern of expression in planta.
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三种植物蔗糖载体在爪蟾卵母细胞中表达的转运特性比较
先前的研究表明,三种植物蔗糖转运体PmSUC1、PmSUC2和AtSUC1的表达模式不同。因此,每个载体可能具有不同的性质,适合于工厂中每个位置的运输功能。为了验证这一假设,通过在非洲爪蟾卵母细胞中表达该蛋白,对每个载体进行了详细的表征。这三种载体在卵母细胞中都具有功能活性,并同时运输蔗糖和麦芽糖,糖引起向内电流,当膜电压越负,电流越高。与PmSUC2相比,AtSUC1和PmSUC1的影响更大。载体对每种糖的亲和力是不同的,与AtSUC1和PmSUC1相比,PmSUC2对蔗糖的Km高10倍。然而,所有三种载体的这些值都依赖于外部pH和膜电压。PmSUC1和PmSUC2在外部pH值为6.5-7.5时实现了最大的糖转运,该值似乎与质子耦合转运机制不一致。然而,用于卵母细胞实验的生理盐水中含有相对高浓度的NaCl,可能会干扰载体蛋白的功能。为了验证这一观点,研究人员在含有较低浓度蔗糖离子的盐水中测量了蔗糖引起的电流。钠离子(而非氯离子)改变了载体的糖转运活性,并且每种载体的作用不同,抑制AtSUC1和PmSUC1,但刺激PmSUC2。讨论了每种载体的特性与它们在卵母细胞中的表达以及它们在植物中的表达模式的关系。
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