MiniVIPER Is a Peptide Tag for Imaging and Translocating Proteins in Cells.,Biochemistry

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MiniVIPER Is a Peptide Tag for Imaging and Translocating Proteins in Cells.
Biochemistry ( IF 2.9 ) Pub Date : 2020-08-07 , DOI: 10.1021/acs.biochem.0c00526
Julia K Doh ₁ , Savannah J Tobin ₁ , Kimberly E Beatty ₁

Affiliation

Microscopy allows researchers to interrogate proteins within a cellular context. To deliver protein-specific contrast, we developed a new class of genetically encoded peptide tags called versatile interacting peptide (VIP) tags. VIP tags deliver a reporter to a target protein via the formation of a heterodimer between the peptide tag and an exogenously added probe peptide. We report herein a new VIP tag named MiniVIPER, which is comprised of a MiniE–MiniR heterodimer. We first demonstrated the selectivity of MiniVIPER by labeling three cellular targets: transferrin receptor 1 (TfR1), histone protein H2B, and the mitochondrial protein TOMM20. We showed that either MiniE or MiniR could serve as the genetically encoded tag. Next, we demonstrated MiniVIPER’s versatility by generating five spectrally distinct probe peptides to label tagged TfR1 on live cells. Lastly, we demonstrated two new applications for VIP tags. First, we used MiniVIPER in combination with another VIP tag, VIPER, to selectively label two different proteins in a single cell (e.g., TfR1 with H2B or TOMM20). Second, we used MiniVIPER to translocate a fluorescent protein to the nucleus through in situ dimerization of mCherry with H2B-mEmerald. In summary, MiniVIPER is a new peptide tag that enables multitarget imaging and artificial dimerization of proteins in cells.

中文翻译：

MiniVIPER是用于成像和转运细胞中蛋白质的肽标签。

显微镜使研究人员可以在细胞环境中探查蛋白质。为了提供蛋白质特异性对比，我们开发了一种新的遗传编码的肽标签，称为通用相互作用肽（VIP）标签。VIP标签通过在肽标签和外源添加的探针肽之间形成异二聚体，将报告基因传递至靶蛋白。我们在此报告了一个名为MiniVIPER的新VIP标签，该标签由MiniE–MiniR异二聚体组成。我们首先通过标记三个细胞靶标来证明MiniVIPER的选择性：转铁蛋白受体1（TfR1），组蛋白H2B和线粒体蛋白TOMM20。我们证明了MiniE或MiniR都可以用作基因编码标签。下一个，我们通过产生五个光谱上不同的探针肽来标记活细胞上标记的TfR1，证明了MiniVIPER的多功能性。最后，我们演示了VIP标签的两个新应用程序。首先，我们将MiniVIPER与另一个VIP标签VIPER结合使用，以选择性标记单个细胞中的两种不同蛋白质（例如，带有H2B或TOMM20的TfR1）。其次，我们使用MiniVIPER通过mCherry与H2B-mEmerald的原位二聚化将荧光蛋白转移到核中。总之，MiniVIPER是一种新的肽标签，可对细胞中的蛋白质进行多目标成像和人工二聚化。我们使用MiniVIPER通过mCherry与H2B-mEmerald的原位二聚化将荧光蛋白转移到核中。总之，MiniVIPER是一种新的肽标签，可对细胞中的蛋白质进行多目标成像和人工二聚化。我们使用MiniVIPER通过mCherry与H2B-mEmerald的原位二聚化将荧光蛋白转移到核中。总之，MiniVIPER是一种新的肽标签，可对细胞中的蛋白质进行多目标成像和人工二聚化。

更新日期：2020-08-25

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