Near-infrared optogenetic pair for protein regulation and spectral multiplexing.
Near-infrared optogenetic pair for protein regulation and spectral multiplexing.
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DOI：
10.1038/nchembio.2343
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发表时间：
2017-06
影响因子：
14.8
通讯作者：
Verkhusha VV
中科院分区：
文献类型：
--
作者：
Redchuk TA;Omelina ES;Chernov KG;Verkhusha VV
多功能光遗传学系统在基础和生物医学研究中的应用需求很高。细菌光敏色素BphP 1与其天然PpsR 2伴侣的近红外光诱导结合有利于与蓝光激活工具同时使用。然而，BphP 1-PpsR 2对的应用受到PpsR 2的大尺寸、多结构域结构和寡聚行为的限制。在这里，我们设计了一个单结构域BphP 1结合伴侣Q-PAS 1，它是三倍小，缺乏寡聚化。我们利用Q-PAS 1的螺旋PAS折叠来开发几种近红外光可控的转录调控系统，使其能够激活或抑制40倍。光诱导的BphP 1-Q-PAS 1相互作用允许染色质表观遗传状态的修改。将BphP 1-Q-PAS 1对与蓝光可激活的基于LOV结构域的系统复用证明了它们的可忽略的光谱串扰。通过将Q-PAS 1和LOV结构域整合到单个光遗传学工具中，我们实现了由近红外光和蓝光独立控制的三向蛋白质靶向，从而证明了Q-PAS 1在光谱复用和多组分系统工程方面的优越性。
Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1–PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix–PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1–Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1–Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.
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影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
14.9
作者：
Müller K;Engesser R;Schulz S;Steinberg T;Tomakidi P;Weber CC;Ulm R;Timmer J;Zurbriggen MD;Weber W
通讯作者：
Weber W
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
64.8
作者：
Bhoo, SH;Davis, SJ;Vierstra, RD
通讯作者：
Vierstra, RD
影响因子：
5.8
作者：
Delorenzi, M;Speed, T
通讯作者：
Speed, T