Chaperone activity of recombinant maize chloroplast protein synthesis elongation factor, EF-Tu.

Damodara Rao, Ivana Momcilovic, Satoru Kobayashi, Eduardo Callegari, Zoran Ristic
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引用次数: 61

Abstract

The protein synthesis elongation factor, EF-Tu, is a protein that carries aminoacyl-tRNA to the A-site of the ribosome during the elongation phase of protein synthesis. In maize (Zea mays L) this protein has been implicated in heat tolerance, and it has been hypothesized that EF-Tu confers heat tolerance by acting as a molecular chaperone and protecting heat-labile proteins from thermal aggregation and inactivation. In this study we investigated the effect of the recombinant precursor of maize EF-Tu (pre-EF-Tu) on thermal aggregation and inactivation of the heat-labile proteins, citrate synthase and malate dehydrogenase. The recombinant pre-EF-Tu was purified from Escherichia coli expressing this protein, and mass spectrometry confirmed that the isolated protein was indeed maize EF-Tu. The purified protein was capable of binding GDP (indicative of protein activity) and was stable at 45 degrees C, the highest temperature used in this study to test this protein for possible chaperone activity. Importantly, the recombinant maize pre-EF-Tu displayed chaperone activity. It protected citrate synthase and malate dehydrogenase from thermal aggregation and inactivation. To our knowledge, this is the first observation of chaperone activity by a plant/eukaryotic pre-EF-Tu protein. The results of this study support the hypothesis that maize EF-Tu plays a role in heat tolerance by acting as a molecular chaperone and protecting chloroplast proteins from thermal aggregation and inactivation.
重组玉米叶绿体蛋白合成伸长因子(EF-Tu)的伴侣活性。
蛋白质合成延伸因子EF-Tu是一种蛋白质,在蛋白质合成的延伸阶段携带氨基酰基trna到核糖体的a位点。在玉米(Zea mays L)中,这种蛋白与耐热性有关,并且已经假设EF-Tu通过作为分子伴侣和保护热不稳定蛋白免受热聚集和失活而赋予耐热性。本研究研究了重组玉米EF-Tu前体(pre-EF-Tu)对热不稳定蛋白、柠檬酸合成酶和苹果酸脱氢酶热聚集和失活的影响。从表达该蛋白的大肠杆菌中纯化得到重组蛋白pre-EF-Tu,质谱分析证实该蛋白确实是玉米EF-Tu。纯化后的蛋白能够结合GDP(指示蛋白活性),并且在45℃下稳定,这是本研究中测试该蛋白可能的伴侣活性的最高温度。重要的是,重组玉米pre-EF-Tu显示出伴侣蛋白活性。它可以防止柠檬酸合酶和苹果酸脱氢酶的热聚集和失活。据我们所知,这是植物/真核生物前ef - tu蛋白首次观察到伴侣蛋白的活性。本研究的结果支持了玉米EF-Tu作为分子伴侣和保护叶绿体蛋白免于热聚集和失活的假设。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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