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科学家开发出可控释放单分子的光遗传学方法
作者:小柯机器人 发布时间:2024/3/13 9:35:33

德国自由柏林大学Helge Ewers团队开发出可控释放单分子的光遗传学方法。2024年3月8日,国际知名学术期刊《自然—方法学》在线发表了这一成果。

研究人员开发了一种在细胞中释放单分子的光遗传学系统。研究人员通过一种可光裂解蛋白将可溶性和跨膜蛋白限制在高尔基体内,并通过短脉冲光释放它们。该方法可根据光剂量将功能蛋白质输送到细胞膜和质膜,其数量与单分子成像相匹配,从而大大简化了对活细胞中任何蛋白质进行单分子显微镜观察的过程。
 
通过将BK和LRRC8/体积调节的阴离子通道输送到质膜,研究人员能够重建离子传导。最后,研究人员通过控制释放被敲除的信号蛋白,在白细胞介素-1的刺激下诱导了T淋巴细胞的NF-kB信号传导。
 
研究人员观察到,光诱导形成的功能性炎症信号复合物仅在活化细胞中,触发核因子kappa-B激酶抑制剂的磷酸化。因此,研究人员开发了一种在单分子水平上重建和研究细胞功能的光遗传学方法。

附:英文原文

Title: An optogenetic method for the controlled release of single molecules

Author: Kashyap, Purba, Bertelli, Sara, Cao, Fakun, Kostritskaia, Yulia, Blank, Fenja, Srikanth, Niranjan A., Schlack-Leigers, Claire, Saleppico, Roberto, Bierhuizen, Dolf, Lu, Xiaocen, Nickel, Walter, Campbell, Robert E., Plested, Andrew J. R., Stauber, Tobias, Taylor, Marcus J., Ewers, Helge

Issue&Volume: 2024-03-08

Abstract: We developed a system for optogenetic release of single molecules in cells. We confined soluble and transmembrane proteins to the Golgi apparatus via a photocleavable protein and released them by short pulses of light. Our method allows for a light dose-dependent delivery of functional proteins to the cytosol and plasma membrane in amounts compatible with single-molecule imaging, greatly simplifying access to single-molecule microscopy of any protein in live cells. We were able to reconstitute ion conductance by delivering BK and LRRC8/volume-regulated anion channels to the plasma membrane. Finally we were able to induce NF-kB signaling in Tlymphoblasts stimulated by interleukin-1 by controlled release of a signaling protein that had been knocked out. We observed light-induced formation of functional inflammatory signaling complexes that triggered phosphorylation of the inhibitor of nuclear factor kappa-B kinase only in activated cells. We thus developed an optogenetic method for the reconstitution and investigation of cellular function at the single-molecule level.

DOI: 10.1038/s41592-024-02204-x

Source: https://www.nature.com/articles/s41592-024-02204-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:47.99
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex

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