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Co-expression of a mammalian accessory trafficking protein enables functional expression of the rat MCT1 monocarboxylate transporter in Saccharomyces cerevisiae

机译:哺乳动物辅助运输蛋白的共表达使大鼠MCT1单羧酸盐转运蛋白在酿酒酵母中功能性表达

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摘要

We have developed a new heterologous expression system for monocarboxylate transporters. The system is based on a Saccharomyces cerevisiae pyk1 mae1 jen1 triple-deletion strain that is auxotrophic for pyruvate and deficient in monocarboxylate uptake. Growth of the yeast cells on ethanol medium supplemented with pyruvate or lactate was dependent on the expression of a suitable monocarboxylate transporter. We have used the system to characterize the functional significance of interactions between the rat MCT1 transporter and its ancillary protein CD147. CD147 was shown to improve trafficking of MCTI to the plasma membrane and its uptake activity. Our results demonstrate a new strategy for the production of properly folded and correctly targeted membrane proteins in a microbial expression system by co-expression of appropriate accessory proteins.
机译:我们已经开发出了一种用于单羧酸盐转运蛋白的新的异源表达系统。该系统基于酿酒酵母pyk1 mae1 jen1三重缺失菌株,该菌株对于丙酮酸是营养缺陷型,单羧酸盐摄取不足。酵母细胞在补充丙酮酸或乳酸的乙醇培养基上的生长取决于合适的单羧酸盐转运蛋白的表达。我们已使用该系统表征大鼠MCT1转运蛋白及其辅助蛋白CD147之间相互作用的功能重要性。已显示CD147可改善MCTI向质膜的运输及其吸收活性。我们的结果证明了通过共表达合适的辅助蛋白在微生物表达系统中产生正确折叠和正确靶向的膜蛋白的新策略。

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