An acidified thermostabilizing mini-peptide derived from the carboxyl extension of the larger isoform of the plant Rubisco activase
An acidified thermostabilizing mini-peptide derived from the carboxyl extension of the larger isoform of the plant Rubisco activase
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一种酸化热稳定小肽，源自植物 Rubisco 激活酶较大亚型的羧基延伸
DOI：
10.1016/j.jbiotec.2015.08.021
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发表时间：
2015
影响因子：
4.1
通讯作者：
Zhurong Zou
中科院分区：
文献类型：
--
作者：
Mengru Zhang;Xujuan Li;Yumei Yang;Zhu Luo;Chang Liu;Ming Gong;Zhurong Zou
热稳定的融合肽伴侣对于蛋白质的热稳定性工程具有重要价值。我们使用蛋白质融合策略评估了拟南芥对应物 (AtRAce) 和植物 Rubisco 激活酶 (RCA) 保守羧基延伸 (RAce) 的酸化衍生物 (mRAce) 在大肠杆菌和酿酒酵母中的热稳定特性。我们使用 AtRAce 和 mRAce 作为来自拟南芥和烟草的不耐热蛋白 RCA2 的融合尾部烟草。 AtRAce 与拟南芥 RCA2 的同源融合以及 AtRAce 与烟草 RCA2 的异源融合均增加了两种蛋白的热稳定性。与 AtRAce 相比，酸化衍生物 mRAce 赋予两种蛋白质更高的热稳定性。此外，mRAce 还增强了麻疯树中其他两种不耐热蛋白的热稳定性：胞质抗坏血酸过氧化物酶 1 (JcAPX1) 和 TATA-box 结合蛋白亚型 1 (JcTBP1)。我们进一步首次报道 JcTBP1 介导酵母体内的耐热性。因此，我们的研究确定了一种 C 端酸性小肽（酸化衍生物 mRAce），其在改善宿主生物体中不耐热蛋白质的热稳定性及其相关耐热性方面具有潜在用途。
Thermostable fusion peptide partners are valuable in engineering thermostability in proteins. We evaluated the Arabidopsis counterpart (AtRAce) and an acidified derivative (mRAce) of the conserved carboxyl extension (RAce) of plant Rubisco activase (RCA) for their thermostabilizing properties inEscherichia coliandSaccharomyces cerevisiaeusing a protein fusion strategy.We used AtRAce and mRAce as fusion tails for the thermolabile protein RCA2 fromArabidopsis thalianaandNicotiana tabacum. The homologous fusion of AtRAce with Arabidopsis RCA2 and the heterologous fusion of AtRAce with tobacco RCA2 increased the thermostability of both proteins. The acidified derivative mRAce conferred greater thermostability upon both proteins as compared with AtRAce. Moreover, mRAce enhanced the thermostability of other two thermolabile proteins fromJatropha curcas: the cytosolic ascorbate peroxidase 1 (JcAPX1) and the TATA-box binding protein isoform 1 (JcTBP1). We further report – for the first time – that JcTBP1 mediates heat tolerancein vivoin yeast. Thus, our study identifies a C-terminal acidic mini-peptide – the acidified derivative mRAce – with potential uses in improving the thermostability of heat-labile proteins and their associated heat tolerance in host organisms.