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Title: 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.

Authors:
ORCiD logo [1]; ORCiD logo [1];  [2];  [1];  [2];  [1];  [1]; ORCiD logo [2];  [3];  [1];  [2]; ORCiD logo [1]
  1. Univ. of Sheffield (United Kingdom)
  2. Washington Univ., St. Louis, MO (United States)
  3. Pennsylvania State Univ., University Park, PA (United States)
Publication Date:
Research Org.:
Washington Univ., St. Louis, MO (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1500070
Grant/Contract Number:  
SC0001035
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Barnett, Samuel F. H., Hitchcock, Andrew, Mandal, Amit K., Vasilev, Cvetelin, Yuen, Jonathan M., Morby, James, Brindley, Amanda A., Niedzwiedzki, Dariusz M., Bryant, Donald A., Cadby, Ashley J., Holten, Dewey, and Hunter, C. Neil. Repurposing a photosynthetic antenna protein as a super-resolution microscopy label. United States: N. p., 2017. Web. doi:10.1038/s41598-017-16834-z.
Barnett, Samuel F. H., Hitchcock, Andrew, Mandal, Amit K., Vasilev, Cvetelin, Yuen, Jonathan M., Morby, James, Brindley, Amanda A., Niedzwiedzki, Dariusz M., Bryant, Donald A., Cadby, Ashley J., Holten, Dewey, & Hunter, C. Neil. Repurposing a photosynthetic antenna protein as a super-resolution microscopy label. United States. doi:10.1038/s41598-017-16834-z.
Barnett, Samuel F. H., Hitchcock, Andrew, Mandal, Amit K., Vasilev, Cvetelin, Yuen, Jonathan M., Morby, James, Brindley, Amanda A., Niedzwiedzki, Dariusz M., Bryant, Donald A., Cadby, Ashley J., Holten, Dewey, and Hunter, C. Neil. Fri . "Repurposing a photosynthetic antenna protein as a super-resolution microscopy label". United States. doi:10.1038/s41598-017-16834-z. https://www.osti.gov/servlets/purl/1500070.
@article{osti_1500070,
title = {Repurposing a photosynthetic antenna protein as a super-resolution microscopy label},
author = {Barnett, Samuel F. H. and Hitchcock, Andrew and Mandal, Amit K. and Vasilev, Cvetelin and Yuen, Jonathan M. and Morby, James and Brindley, Amanda A. and Niedzwiedzki, Dariusz M. and Bryant, Donald A. and Cadby, Ashley J. and Holten, Dewey and Hunter, C. Neil},
abstractNote = {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.},
doi = {10.1038/s41598-017-16834-z},
journal = {Scientific Reports},
number = 1,
volume = 7,
place = {United States},
year = {2017},
month = {12}
}

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