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首页> 外文期刊>American Journal of Physiology >Functional expression of heterologous proteins in yeast: insights into Ca2+ signaling and Ca2+-transporting ATPases.
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Functional expression of heterologous proteins in yeast: insights into Ca2+ signaling and Ca2+-transporting ATPases.

机译：酵母中异源蛋白质的功能表达：洞悉Ca2 +信号传导和Ca2 +转运ATPase。

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摘要

The baker's yeast Saccharomyces cerevisiae is a well-developed, versatile, and widely used model organism. It offers a compact and fully sequenced genome, tractable genetics, simple and inexpensive culturing conditions, and, importantly, a conservation of basic cellular machinery and signal transducing pathways with higher eukaryotes. In this review, we describe recent technical advances in the heterologous expression of proteins in yeast and illustrate their application to the study of the Ca(2+) homeostasis machinery, with particular emphasis on Ca(2+)-transporting ATPases. Putative Ca(2+)-ATPases in the newly sequenced genomes of organisms such as parasites, plants, and vertebrates have been investigated by functional complementation of an engineered yeast strain lacking endogenous Ca(2+) pumps. High-throughput screens of mutant phenotypes to identify side chains critical for ion transport and selectivity have facilitated structure-function analysis, and genomewide approaches may be used to dissect cellular pathways involved in Ca(2+) transport and trafficking. The utility of the yeast system is demonstrated by rapid advances in the study of the emerging family of Golgi/secretory pathway Ca(2+),Mn(2+)-ATPases (SPCA). Functional expression of human SPCA1 in yeast has provided insight into the physiology, novel biochemical characteristics, and subcellular localization of this pump. Haploinsufficiency of SPCA1 leads to Hailey-Hailey disease (HDD), a debilitating blistering disorder of the skin. Missense mutations, identified in patients with HHD, may be conveniently assessed in yeast for loss-of-function phenotypes associated with the disease.
机译：面包酵母酿酒酵母是一种成熟的，用途广泛且用途广泛的模型生物。它提供了紧凑而完整的基因组序列，易于控制的遗传学，简单且廉价的培养条件，并且重要的是，它保留了基本的细胞机器以及具有较高真核生物的信号转导途径。在这篇综述中，我们描述了酵母中蛋白质异源表达的最新技术进展，并阐明了它们在Ca（2+）稳态机制研究中的应用，尤其着重于Ca（2+）转运ATPase。通过对缺乏内源性Ca（2+）泵的工程酵母菌株进行功能互补，研究了诸如寄生虫，植物和脊椎动物等生物的新测序基因组中的推定Ca（2 +）-ATPase。高通量的突变体表型筛选，以识别对离子运输和选择性至关重要的侧链，已经促进了结构功能分析，并且全基因组方法可用于剖析参与Ca（2+）运输和运输的细胞途径。通过对高尔基体/分泌途径Ca（2 +），Mn（2 +）-ATPases（SPCA）的新兴家族研究的快速进展证明了酵母系统的实用性。人SPCA1在酵母中的功能表达提供了对该泵的生理学，新的生化特征和亚细胞定位的了解。 SPCA1的单倍剂量不足会导致Hailey-Hailey病（HDD），这是一种使人虚弱的皮肤水疱病。在HHD患者中发现的错义突变可以方便地在酵母中评估与该疾病相关的功能丧失表型。

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