Fluorescent Labelling of Cell Surface Proteins on a Solid Support,Israel Journal of Chemistry

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Fluorescent Labelling of Cell Surface Proteins on a Solid Support
Israel Journal of Chemistry ( IF 2.3 ) Pub Date : 2021-04-19 , DOI: 10.1002/ijch.202100028
Sean Adler ₁ , Leila Motiei ₁ , Naama Mankovski ₁ , Hagai Cohen ₂ , David Margulies ₁

Affiliation

A method for decorating bacteria with synthetic receptors while they are attached to a solid support is described. This approach involves two steps. First, E. coli expressing a His‐tagged outer membrane protein C (His‐OmpC) are attached to a monolayer of DNA duplexes appended with a tri‐nitrilotriacetic acid (tri‐NTA) group. The remaining His‐tags, which are not involved in the interaction with the monolayer, can then be modified with additional tri‐NTA‐appended duplexes that bear different functionalities. By labelling the surface‐bound bacteria with an oligodeoxynucleotide (ODN) appended with a fluorescent dye and by turning off its emission using a complementary, quencher‐bearing strand, we demonstrate the versatility of this approach, as well as a means to modify cell membrane proteins on a solid support using simple self‐assembly processes.

中文翻译：

固体支持物上的细胞表面蛋白的荧光标记

描述了一种在合成受体附着于固相支持物时用细菌修饰细菌的方法。此方法涉及两个步骤。首先，表达带有His标签的外膜蛋白C（His-OmpC）的大肠杆菌附着在DNA双链体的单分子层上，并附加了三硝基三乙酸（tri-NTA）基团。其余的不包含在与单层相互作用中的His-tags，可以用带有不同功能的附加的tri-NTA-附加的双链体进行修饰。通过用寡聚脱氧核苷酸（ODN）标记荧光染料并通过使用带有淬灭剂的互补链关闭其发射，对表面结合的细菌进行标记，我们证明了这种方法的多功能性以及修饰细胞膜的方法使用简单的自组装过程将蛋白质固定在固体支持物上。

更新日期：2021-04-23

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