Sulfonated red and far-red rhodamines to visualize SNAP- and Halo-tagged cell surface proteins.
Sulfonated red and far-red rhodamines to visualize SNAP- and Halo-tagged cell surface proteins.
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磺化红和远红罗丹明用于可视化SNAP和Halo标记的细胞表面蛋白。
DOI：
10.1039/d1ob02216d
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发表时间：
2022-08-03
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
--
作者：
渗透膜的能力是化学生物学探针的一个关键特征，它定义了它们对特定应用的适用性。在这里，我们报告磺化罗丹明，赋予氧杂蒽染料与细胞的不渗透性的表面蛋白质的分析。我们融合带电的磺酸盐红色和远红染料，以获得Sulfo 549和Sulfo 646，分别，并进一步将这些连接到苄基鸟嘌呤和choloralkane底物SNAP标签和卤素标签标记。磺化罗丹明共轭荧光团保持理想的物理性能，如亮度和光稳定性。虽然具有核定位的SNAP标签的转染细胞保持未标记，但细胞外暴露的标签可以清楚地可视化。通过与可渗透的罗丹明的多路复用，我们能够区分细胞外和细胞内SNAP-和Halo-标签，包括那些安装在胰高血糖素样肽-1受体，一个典型的B G类蛋白偶联受体。Sulfo 549和Sulfo 646还标记了来自诱导多能干细胞（iPSC）的转染神经元，允许轴突膜的STED纳米显微镜。总之，这项工作提供了一个新的途径，使染料不可渗透的独家细胞外可视化通过自我标记的蛋白质标签。我们预计，Sulfo 549，Sulfo 646和它们的同类物将是有用的细胞生物学应用中的细胞内位点的标记干扰准确的表面蛋白分析。磺化罗丹明染料允许SNAP-和Halo-标签标记细胞表面蛋白融合。远红外版本可用于STED纳米显微镜。
The (in)ability to permeate membranes is a key feature of chemical biology probes that defines their suitability for specific applications. Here we report sulfonated rhodamines that endow xanthene dyes with cellular impermeability for analysis of surface proteins. We fuse charged sulfonates to red and far-red dyes to obtain Sulfo549 and Sulfo646, respectively, and further link these to benzylguanine and choloralkane substrates for SNAP-tag and Halo-tag labelling. Sulfonated rhodamine-conjugated fluorophores maintain desirable photophysical properties, such as brightness and photostability. While transfected cells with a nuclear localized SNAP-tag remain unlabelled, extracellular exposed tags can be cleanly visualized. By multiplexing with a permeable rhodamine, we are able to differentiate extra- and intracellular SNAP- and Halo-tags, including those installed on the glucagon-like peptide-1 receptor, a prototypical class B G protein-coupled receptor. Sulfo549 and Sulfo646 also labelled transfected neurons derived from induced pluripotent stem cells (iPSCs), allowing STED nanoscopy of the axonal membrane. Together, this work provides a new avenue for rendering dyes impermeable for exclusive extracellular visualization via self-labelling protein tags. We anticipate that Sulfo549, Sulfo646 and their congeners will be useful for a number of cell biology applications where labelling of intracellular sites interferes with accurate surface protein analysis. Sulfonated rhodamine dyes allow SNAP- and Halo-tag labelling of cell surface protein fusions. A far-red version can be used for STED nanoscopy.
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影响因子：
8
作者：
Grimm JB;Xie L;Casler JC;Patel R;Tkachuk AN;Falco N;Choi H;Lippincott-Schwartz J;Brown TA;Glick BS;Liu Z;Lavis LD
通讯作者：
Lavis LD
影响因子：
48
作者：
Asher WB;Geggier P;Holsey MD;Gilmore GT;Pati AK;Meszaros J;Terry DS;Mathiasen S;Kaliszewski MJ;McCauley MD;Govindaraju A;Zhou Z;Harikumar KG;Jaqaman K;Miller LJ;Smith AW;Blanchard SC;Javitch JA
通讯作者：
Javitch JA
影响因子：
4.3
作者：
Kolmakov, Kirill;Belov, Vladimir N.;Hell, Stefan W.
通讯作者：
Hell, Stefan W.
影响因子：
7.7
作者：
Gutzeit, Vanessa A.;Thibado, Jordana;Levitz, Joshua
通讯作者：
Levitz, Joshua
影响因子：
8.8
作者：
Lee, Joon;Munguba, Hermany;Levitz, Joshua
通讯作者：
Levitz, Joshua