Repurposing a photosynthetic antenna protein as a super-resolution microscopy label

Abstract

Techniques such as Stochastic Optical Reconstruction Microscopy (STORM) and Structured Illumination Microscopy (SIM) have increased the achievable resolution of optical imaging, but few fluorescent proteins are suitable for super-resolution microscopy, particularly in the far-red and near-infrared emission range. Here we demonstrate the applicability of CpcA, a subunit of the photosynthetic antenna complex in cyanobacteria, for STORM and SIM imaging. The periodicity and width of fabricated nanoarrays of CpcA, with a covalently attached phycoerythrobilin (PEB) or phycocyanobilin (PCB) chromophore, matched the lines in reconstructed STORM images. SIM and STORM reconstructions of Escherichia coli cells harbouring CpcA-labelled cytochrome bd 1 ubiquinol oxidase in the cytoplasmic membrane show that CpcA-PEB and CpcA-PCB are suitable for super-resolution imaging in vivo. The stability, ease of production, small size and brightness of CpcA-PEB and CpcA-PCB demonstrate the potential of this largely unexplored protein family as novel probes for super-resolution microscopy.

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Item Type:	Article
Authors/Creators:	Barnett, S.F.H. https://orcid.org/0000-0002-3696-3509 Hitchcock, A. https://orcid.org/0000-0001-6572-434X Mandal, A.K. Vasilev, C. Yuen, J.M. Morby, J. Brindley, A.A. Niedzwiedzki, D.M. https://orcid.org/0000-0002-1976-9296 Bryant, D.A. Cadby, A.J. Holten, D. Hunter, C.N. https://orcid.org/0000-0003-2533-9783
Copyright, Publisher and Additional Information:	© 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Keywords:	Biological fluorescence; Fluorescence imaging
Dates:	Accepted: 16 November 2017 Published (online): 1 December 2017 Published: 23 March 2017
Institution:	The University of Sheffield
Academic Units:	The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) > Department of Molecular Biology and Biotechnology (Sheffield)
Funding Information:	Funder Grant number WASHINGTON UNIVERSITY IN ST. LOUIS WU-HT-10-12 BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL (BBSRC) BB/M000265/1
Depositing User:	Symplectic Sheffield
Date Deposited:	14 Dec 2017 15:38
Last Modified:	21 Dec 2023 10:39
Status:	Published
Publisher:	Nature Publishing Group
Refereed:	Yes
Identification Number:	10.1038/s41598-017-16834-z
Related URLs:	Author
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:124984

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Repurposing a photosynthetic antenna protein as a super-resolution microscopy label

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