Cell wall trapping of autocrine peptides for human G-protein-coupled receptors on the yeast cell surface.

Cell wall trapping of autocrine peptides for human G-protein-coupled receptors on the yeast cell surface.
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DOI:
10.1371/journal.pone.0037136
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Kondo A
Kondo A
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Ishii J;Yoshimoto N;Tatematsu K;Kuroda S;Ogino C;Fukuda H;Kondo A

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G蛋白偶联受体(GPCRs)调节多种生理过程,是药物发现的重要药物靶点。在这里,我们描述了一种基于酵母细胞表面展示技术的独特概念,以选择性地跟踪符合条件的具有人类GPCRs(细胞壁自分泌多肽捕获)策略的激活性多肽(CWTrAP)。在我们的策略中,单个重组酵母细胞能够通过表达融合到锚定基序的候选肽来报告人类GPCRs的自分泌阳性活性。在表达和激活之后,酵母细胞将自分泌肽捕获到细胞壁上。因为捕获的多肽不能扩散,所以它们不会对周围表达目标人类GPCR和非信号肽的酵母细胞产生影响。因此,单个酵母细胞可以组装自主信号复合体,并允许对酵母群体进行单细胞筛选。我们的策略可能被应用于寻找对靶向人GPCRs具有激动性活性的合格多肽。
G-protein-coupled receptors (GPCRs) regulate a wide variety of physiological processes and are important pharmaceutical targets for drug discovery. Here, we describe a unique concept based on yeast cell-surface display technology to selectively track eligible peptides with agonistic activity for human GPCRs (Cell Wall Trapping of Autocrine Peptides (CWTrAP) strategy). In our strategy, individual recombinant yeast cells are able to report autocrine-positive activity for human GPCRs by expressing a candidate peptide fused to an anchoring motif. Following expression and activation, yeast cells trap autocrine peptides onto their cell walls. Because captured peptides are incapable of diffusion, they have no impact on surrounding yeast cells that express the target human GPCR and non-signaling peptides. Therefore, individual yeast cells can assemble the autonomous signaling complex and allow single-cell screening of a yeast population. Our strategy may be applied to identify eligible peptides with agonistic activity for target human GPCRs.
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