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Sci Rep. 2017 Dec 1;7(1):16807. doi: 10.1038/s41598-017-16834-z.

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

Author information

1
Department of Molecular Biology and Biotechnology, Firth Court, Western Bank, Sheffield, S10 2TN, UK.
2
Department of Chemistry, Washington University, St. Louis, MO, 63130, USA.
3
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania, 16802, USA.
4
Department of Physics and Astronomy, Hicks Building, Hounsfield Road, Sheffield, S3 7RH, UK.
5
Department of Molecular Biology and Biotechnology, Firth Court, Western Bank, Sheffield, S10 2TN, UK. c.n.hunter@sheffield.ac.uk.

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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