Fluorescence energy transfer in the bi-fluorescent S-layer tandem fusion protein ECFP-SgsE-YFP

Journal of Structural Biology
Birgit KainzJ L Toca-Herrera

Abstract

This work reports for the first time on the fabrication of a bi-functional S-layer tandem fusion protein which is able to self-assemble on solid supports without losing its functionality. Two variants of the green fluorescent protein (GFP) were genetically combined with a self-assembly system having the remarkable opportunity to interact with each other and act as functional nanopatterning biocoating. The S-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a was fused with the cyan ECFP donor protein at the SgsE N-terminus and with the yellow YFP acceptor protein at the C-terminus. The fluorescence energy transfer was studied with spectrofluorimetry, confocal microscopy and flow cytometry, whilst protein self-assembly (on silicon dioxide particles) and structural investigations were carried out with atomic force microscopy (AFM). The fluorescence resonance energy transfer efficiency of reassembled SgsE tandem protein was 20.0 ± 6.1% which is almost the same transfer efficiency shown in solution (19.6 ± 0.1%). This work shows that bi-fluorescent S-layer fusion proteins self-assemble on silica particles retaining their fluorescent properties.

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Citations

Oct 5, 2013·The Journal of Chemical Physics·Ainhoa LejardiJosé L Toca-Herrera
Jun 24, 2011·Current Opinion in Biotechnology·Nicola IlkUwe B Sleytr
Feb 4, 2014·FEMS Microbiology Reviews·Uwe B SleytrDietmar Pum
Mar 31, 2015·Acta Biomaterialia·Bernhard Schuster, Uwe B Sleytr

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