VIPER is a genetically encoded peptide tag for fluorescence and electron microscopy

Julia K. Doh, Jonathan D. White, Hannah K. Zane, Young Hwan Chang, Claudia Lopez, Caroline Enns, Kimberly Beatty

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Many discoveries in cell biology rely on making specific proteins visible within their native cellular environment. There are various genetically encoded tags, such as fluorescent proteins, developed for fluorescence microscopy (FM). However, there are almost no genetically encoded tags that enable cellular proteins to be observed by both FM and electron microscopy (EM). Herein, we describe a technology for labeling proteins with diverse chemical reporters, including bright organic fluorophores for FM and electron-dense nanoparticles for EM. Our technology uses versatile interacting peptide (VIP) tags, a class of genetically encoded tag. We present VIPER, which consists of a coiled-coil heterodimer formed between the genetic tag, CoilE, and a probe-labeled peptide, CoilR. Using confocal FM, we demonstrate that VIPER can be used to highlight subcellular structures or to image receptor-mediated iron uptake. Additionally, we used VIPER to image the iron uptake machinery by correlative light and EM (CLEM). VIPER compared favorably with immunolabeling for imaging proteins by CLEM, and is an enabling technology for protein targets that cannot be immunolabeled. VIPER is a versatile peptide tag that can be used to label and track proteins with diverse chemical reporters observable by both FM and EM instrumentation.

Original languageEnglish (US)
Pages (from-to)12961-12966
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number51
DOIs
StatePublished - Dec 18 2018

Keywords

  • Biochemistry
  • Coiled coil
  • Electron microscopy
  • Fluorescence microscopy
  • Hemical biology

ASJC Scopus subject areas

  • General

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