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化学诱导异源三聚体系统的理性设计与实现
作者:小柯机器人 发布时间:2020/8/5 15:58:05

约翰·霍普金斯大学医学院的Takanari Inoue课题组和日本理研所(RIKEN)的Takashi Umehara课题组合作在研究中取得进展。他们的最新研究提出了理性设计和实现化学诱导异源三聚体的方法。 2020年8月3日,《自然—方法学》发表了这一成果。

研究团队通过理性分裂FRB和FKBP,开发了化学诱导三聚(CIT)的方法。细胞和结构数据集显示,雷帕霉素分别有效地将FRB或FKBP拆分对与全长FKBP或FRB进行了三聚。CIT迅速诱导三细胞器(内质网-细胞质膜-高尔基体)的连接,并在选择的膜接触位点扰动特定膜脂质。在CID的现有基础上,科研人员通过赋予一附加条件开发了CIT。CIT扩展了单个活细胞中的操纵类型,以解决其他方式难以解决的细胞生物学问题,并为将来的合成生物学应用提供工程化细胞功能的工具。

据介绍,化学诱导二聚化(CID)使用小分子诱导两种不同蛋白质的结合。CID工具,例如FK506结合蛋白– FKBP –雷帕霉素结合–(FKBP–FRB)–雷帕霉素系统,已被广泛用于探测细胞内外分子事件。尽管已有多种CID工具,但由于设计可同时以高亲和力和特异性结合三种蛋白质的化学药品的固有挑战,化学诱导三聚(CIT)不存在。

附:英文原文

Title: Rational design and implementation of a chemically inducible heterotrimerization system

Author: Helen D. Wu, Masaki Kikuchi, Onur Dagliyan, Adam K. Aragaki, Hideki Nakamura, Nikolay V. Dokholyan, Takashi Umehara, Takanari Inoue

Issue&Volume: 2020-08-03

Abstract: Chemically inducible dimerization (CID) uses a small molecule to induce binding of two different proteins. CID tools such as the FK506-binding protein–FKBP–rapamycin-binding– (FKBP–FRB)–rapamycin system have been widely used to probe molecular events inside and outside cells. While various CID tools are available, chemically inducible trimerization (CIT) does not exist, due to inherent challenges in designing a chemical that simultaneously binds three proteins with high affinity and specificity. Here, we developed CIT by rationally splitting FRB and FKBP. Cellular and structural datasets showed efficient trimerization of split pairs of FRB or FKBP with full-length FKBP or FRB, respectively, by rapamycin. CIT rapidly induced tri-organellar junctions and perturbed intended membrane lipids exclusively at select membrane contact sites. By conferring one additional condition to what is achievable with CID, CIT expands the types of manipulation in single live cells to address cell biology questions otherwise intractable and engineer cell functions for future synthetic biology applications.

DOI: 10.1038/s41592-020-0913-x

Source: https://www.nature.com/articles/s41592-020-0913-x

期刊信息

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex