Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes,RSC Chemical Biology

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Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes
RSC Chemical Biology ( IF 4.2 ) Pub Date : 2021-07-16 , DOI: 10.1039/d1cb00126d
Georgina C Gavins ₁ , Katharina Gröger ₁ , Marc Reimann ₁ , Michael D Bartoschek ₂ , Sebastian Bultmann ₂ , Oliver Seitz ₁

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Templated chemistry offers the prospect of addressing specificity challenges occurring in bioconjugation reactions. Here, we show two peptide-templated amide-bond forming reactions that enable the concurrent labelling of two different membrane proteins with two different peptide nucleic acid (PNA) barcodes. The reaction system is based on the mutually selective coiled coil interaction between two thioester-linked PNA–peptide conjugates and two cysteine peptides serving as genetically encoded peptide tags. Orthogonal coiled coil templated covalent labelling is highly specific, quantitative and proceeds within a minute. To demonstrate the usefulness, we evaluated receptor internalisation of two membranous receptors EGFR (epidermal growth factor) and ErbB2 (epidermal growth factor receptor 2) by first staining PNA-tagged proteins with fluorophore–DNA conjugates and then erasing signals from non-internalized receptors via toehold-mediated strand displacement.

中文翻译：

正交卷曲线圈能够使用肽核酸条形码快速共价标记两种不同的膜蛋白

模板化学为解决生物偶联反应中发生的特异性挑战提供了前景。在这里，我们展示了两种以肽为模板的酰胺键形成反应，这些反应能够同时标记两种不同的膜蛋白与两种不同的肽核酸 ( PNA ) 条形码。该反应系统基于两个硫酯连接的PNA之间的相互选择性卷曲线圈相互作用– 肽缀合物和两个半胱氨酸肽作为基因编码的肽标签。正交卷曲线圈模板化共价标记具有高度特异性、定量性，并且在一分钟内即可完成。为了证明其实用性，我们通过首先用荧光团-DNA 偶联物染色PNA标记的蛋白质，然后从非内化受体通过立足点介导的链置换。

更新日期：2021-07-22

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