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

By Samuel F. H. Barnett, A. Hitchcock, A. K. Mandal, Cvetelin Vasilev, J. Yuen, J. Morby, Amanda A Brindley, D. Niedzwiedzki, Donald A. Bryant, Ashley J. Cadby and 2 more. 2017

5 citations recorded by the source corpus.

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 bd1 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.

Subfields: Biophysics, Molecular Biology, Structural Biology

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In Molecular Biology

1. Minkyung Baek
2. Ivan V. Anishchenko
3. Frank DiMaio
4. A. Kang
5. Jue Wang
6. Ryan McHugh
7. A. Bera
8. David Baker

In Structural Biology

1. Frank DiMaio

Citations in the corpus

• Generalized Biomolecular Modeling and Design with RoseTTAFold All-Atom (2023, positive): “Conformationally restricted bilins typically display higher fluorescence yields; absolute fluorescent yields for the BIL_ C11, BIL_H4, and BIL_F9 designs are 38, 11, and 25%, respectively, based on an earlier determination of the absolute fl uorescence quantum yield for CpcA-PEB of 67% ( 63 ) (fig.”