Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy

Overview

Journal Bio Protoc

Specialty Biology

Date 2021 Mar 3

PMID 33654913

Citations 3

Authors

Julia K Doh

Young Hwan Chang

Caroline A Enns

Claudia S Lopez

Kimberly E Beatty

Affiliations

Soon will be listed here.

Abstract

Advances in fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) offer unprecedented opportunities for studying diverse proteins and nanostructures involved in fundamental cell biology. It is now possible to visualize and quantify the spatial organization of cellular proteins and other macromolecules by FM, EM, and CLEM. However, tagging and tracking cellular proteins across size scales is restricted by the scarcity of methods for attaching appropriate reporter chemistries to target proteins. Namely, there are few genetic tags compatible with EM. To overcome these issues we developed Versatile Interacting Peptide (VIP) tags, genetically-encoded peptide tags that can be used to image proteins by fluorescence and EM. VIPER, a VIP tag, can be used to label cellular proteins with bright, photo-stable fluorophores for FM or electron-dense nanoparticles for EM. In this Bio-Protocol, we provide an instructional guide for implementing VIPER for imaging a cell-surface receptor by CLEM. This protocol is complemented by two other Bio-Protocols outlining the use of VIPER ( Doh , 2019a and 2019b).

Citing Articles

Orthogonal Versatile Interacting Peptide Tags for Imaging Cellular Proteins.

Suyama A, Devlin K, Macias-Contreras M, Doh J, Shinde U, Beatty K Biochemistry. 2023; 62(11):1735-1743.

PMID: 37167569 PMC: 10249344. DOI: 10.1021/acs.biochem.2c00712.

Generation of CoilR Probe Peptides for VIPER-labeling of Cellular Proteins.

Doh J, Tobin S, Beatty K Bio Protoc. 2021; 9(21):e3412.

PMID: 33654912 PMC: 7853930. DOI: 10.21769/BioProtoc.3412.

Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy.

Doh J, Enns C, Beatty K Bio Protoc. 2020; 9(21).

PMID: 32665966 PMC: 7360171. DOI: 10.21769/bioprotoc.3413.

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