Covalent Protein Labeling by SpyTag-SpyCatcher in Fixed Cells for Super-Resolution Microscopy

Chembiochem : a European Journal of Chemical Biology
Veronica PessinoBo Huang

Abstract

Labeling proteins with high specificity and efficiency is a fundamental prerequisite for microscopic visualization of subcellular protein structures and interactions. Although the comparatively small size of epitope tags makes them less perturbative to fusion proteins, they require the use of large antibodies that often limit probe accessibility and effective resolution. Here we use the covalent SpyTag-SpyCatcher system as an epitope-like tag for fluorescent labeling of intracellular proteins in fixed cells for both conventional and super-resolution microscopy. We also applied this method to endogenous proteins by gene editing, demonstrating its high labeling efficiency and capability for isoform-specific labeling.

References

Jun 7, 2008·ACS Chemical Biology·Georgyi V LosKeith V Wood
Nov 26, 2010·Nature·Pakorn KanchanawongClare M Waterman
Dec 15, 2010·Neuron·Adish DaniXiaowei Zhuang
Mar 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Bijan ZakeriMark Howarth
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jul 27, 2012·Annual Review of Cell and Developmental Biology·Frances M Brodsky
Sep 29, 2012·Nature Methods·Felipe OpazoSilvio O Rizzoli
Feb 11, 2014·Current Protocols in Protein Science·Nelson B Cole
Jul 24, 2015·Angewandte Chemie·Ralph WienekeRobert Tampé
Jul 28, 2015·Chemistry & Biology·Claire N BedbrookViviana Gradinaru
Jan 21, 2016·Proceedings of the National Academy of Sciences of the United States of America·Gianluca VeggianiMark Howarth
Mar 19, 2016·Nature Communications·Daichi KamiyamaBo Huang
Jun 9, 2016·Proceedings of the National Academy of Sciences of the United States of America·Manuel D LeonettiBo Huang
Nov 18, 2016·Chemical Communications : Chem Comm·Hyojin MoonSebyung Kang
Feb 15, 2017·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Christos KarathanasisMike Heilemann

❮ Previous
Next ❯

Citations

Dec 12, 2017·Protein Engineering, Design & Selection : PEDS·M HinrichsenL Regan
Oct 13, 2017·Angewandte Chemie·Anthony H KeebleMark Howarth
Dec 6, 2019·Biochemical Society Transactions·Rebecca E A GwytherHarley L Worthy
Apr 17, 2019·Nature Communications·Irsyad N A Khairil AnuarMark Howarth
May 6, 2019·International Journal of Molecular Sciences·Daniel HatlemJack C Leo
Apr 18, 2018·Nature Communications·Wenwei MaEnrico Ferrari
Feb 9, 2021·Chemical Science·Anthony H Keeble, Mark Howarth
Feb 8, 2021·International Journal of Biological Macromolecules·Yoonji BaeSebyung Kang
Jun 8, 2021·The Journal of Cell Biology·Anne PipathsoukOrion D Weiner
Sep 18, 2018·Chemical Reviews·Honglin Li, Joshua C Vaughan

❮ Previous
Next ❯

Related Concepts

Related Feeds

BioHub - Researcher Network

The Chan-Zuckerberg Biohub aims to support the fundamental research and develop the technologies that will enable physicians to cure, prevent, or manage all diseases in our childrens' lifetimes. The CZ Biohub brings together researchers from UC Berkeley, Stanford, and UCSF. Find the latest research from the CZ Biohub researcher network here.