Miller A J, Smith S J, Theodoulou F L
Biochemistry and Physiology Department, Rothamsted Experimental Station, AFRC-IACR, Harpenden, Hertfordshire, UK.
Symp Soc Exp Biol. 1994;48:167-77.
The Xenopus oocyte has become a convenient and robust system for the expression of many different animal transport proteins and it has now been demonstrated that oocytes can also translate, process and target plant membrane proteins. Here we review the background to this expression system and discuss how it can be used to express plant transporters. A H+/hexose cotransporter (STP1) from Arabidopsis thaliana has been successfully expressed in oocytes and further characterization has shown the transporter to have properties similar to the same transporter expressed in yeast. However, the expression of H(+)-coupled transporters may present problems for oocyte intracellular pH regulation, because the influx of protons during their operation may acidify the cell. To investigate this possibility, proton-selective microelectrodes have been used to measure intracellular pH during symport of H+ and hexose in oocytes expressing STP1. These measurements showed that oocyte cytosolic pH is unaltered during H+/glucose cotransport, but is sensitive to changes in external pH.
非洲爪蟾卵母细胞已成为表达多种不同动物转运蛋白的便捷且强大的系统,目前已有研究表明卵母细胞还能翻译、加工并靶向定位植物膜蛋白。在此,我们回顾该表达系统的背景,并讨论如何利用它来表达植物转运蛋白。拟南芥的一种H⁺/己糖共转运蛋白(STP1)已在卵母细胞中成功表达,进一步的特性分析表明该转运蛋白具有与在酵母中表达的相同转运蛋白相似的特性。然而,H⁺偶联转运蛋白的表达可能会给卵母细胞的细胞内pH调节带来问题,因为在其运作过程中质子的流入可能会使细胞酸化。为了研究这种可能性,已使用质子选择性微电极来测量表达STP1的卵母细胞在H⁺和己糖同向转运过程中的细胞内pH。这些测量结果表明,在H⁺/葡萄糖共转运过程中,卵母细胞的胞质pH没有变化,但对外部pH的变化敏感。