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利用HaloTag技术对活细胞中蛋白质进行位点特异性标记的先进荧光聚合物探针

Advanced Fluorescent Polymer Probes for the Site-Specific Labeling of Proteins in Live Cells Using the HaloTag Technology.

作者信息

Berki Thomas, Bakunts Anush, Duret Damien, Fabre Laura, Ladavière Catherine, Orsi Andrea, Charreyre Marie-Thérèse, Raimondi Andrea, van Anken Eelco, Favier Arnaud

机构信息

Univ Lyon, Université Lyon 1, INSA de Lyon, CNRS, Laboratoire Ingénierie des Matériaux Polymères, UMR5223, F-69621 Villeurbanne, France.

Univ Lyon, Ens de Lyon, CNRS, Laboratoire Joliot-Curie, USR3010, F-69364 Lyon, France.

出版信息

ACS Omega. 2019 Jul 30;4(7):12841-12847. doi: 10.1021/acsomega.9b01643. eCollection 2019 Jul 31.

Abstract

We report the site-specific and covalent bioconjugation of fluorescent polymer chains to proteins in live cells using the HaloTag technology. Polymer chains bearing a Halo-ligand precisely located at their α-chain-end were synthesized in a controlled manner owing to the RAFT polymerization process. They were labeled in lateral position by several organic fluorophores such as AlexaFluor 647. The resulting Halo-ligand polymer probe was finally shown to selectively recognize and label HaloTag proteins present at the membrane of live cells using confocal fluorescence microscopy. Such a polymer bioconjugation approach holds great promises for various applications ranging from cell imaging to cell surface functionalization.

摘要

我们报告了使用HaloTag技术在活细胞中将荧光聚合物链与蛋白质进行位点特异性和共价生物共轭。由于可逆加成-断裂链转移(RAFT)聚合过程,精确位于其α链末端的带有Halo配体的聚合物链得以可控合成。它们通过几种有机荧光团(如AlexaFluor 647)在侧向位置进行标记。最终,使用共聚焦荧光显微镜显示,所得的Halo配体聚合物探针能够选择性地识别并标记活细胞膜上存在的HaloTag蛋白。这种聚合物生物共轭方法在从细胞成像到细胞表面功能化等各种应用中具有巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec91/6682114/7a8dec7d18f0/ao-2019-01643t_0007.jpg

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