Engineering a rigid protein tunnel for biomolecular detection.

Journal of the American Chemical Society
Mohammad M MohammadLiviu Movileanu

Abstract

One intimidating challenge in protein nanopore-based technologies is designing robust protein scaffolds that remain functionally intact under a broad spectrum of detection conditions. Here, we show that an extensively engineered bacterial ferric hydroxamate uptake component A (FhuA), a β-barrel membrane protein, functions as a robust protein tunnel for the sampling of biomolecular events. The key implementation in this work was the coupling of direct genetic engineering with a refolding approach to produce an unusually stable protein nanopore. More importantly, this nanostructure maintained its stability under many experimental circumstances, some of which, including low ion concentration and highly acidic aqueous phase, are normally employed to gate, destabilize, or unfold β-barrel membrane proteins. To demonstrate these advantageous traits, we show that the engineered FhuA-based protein nanopore functioned as a sensing element for examining the proteolytic activity of an enzyme at highly acidic pH and for determining the kinetics of protein-DNA aptamer interactions at physiological salt concentration.

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Citations

Aug 8, 2013·Journal of the American Chemical Society·Misha SoskineGiovanni Maglia
Mar 1, 2013·ACS Nano·David J NiedzwieckiLiviu Movileanu
Feb 8, 2014·PloS One·Besnik Krasniqi, Jeremy S Lee
Jul 23, 2015·Proceedings of the National Academy of Sciences of the United States of America·James A StapletonVikas Nanda
Oct 13, 2015·Biochimica Et Biophysica Acta·Aaron J WolfeLiviu Movileanu
Oct 17, 2015·Nano Letters·Hajar Mamad-HemouchJuan Pelta
Jul 25, 2012·Biochimica Et Biophysica Acta·Jiaming LiuLiviu Movileanu
Dec 19, 2012·Biochimica Et Biophysica Acta·Noriko TomitaLiviu Movileanu
Apr 14, 2015·The Journal of Physical Chemistry. B·Emma Letizia BonomeMauro Chinappi
Dec 4, 2012·Biophysical Journal·David J NiedzwieckiLiviu Movileanu
Jul 6, 2016·ACS Chemical Biology·Mohammad M MohammadLiviu Movileanu
Oct 21, 2014·Nanoscale·L J SteinbockA Radenovic
Jun 21, 2017·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Kherim WillemsGiovanni Maglia
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Nov 12, 2013·Angewandte Chemie·Yi-Lun YingYi-Tao Long
Sep 14, 2019·Current Protocols in Protein Science·Aaron J WolfeLiviu Movileanu
Jul 6, 2014·The Analyst·Yi-Lun YingYi-Tao Long
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Jul 22, 2018·Nature Communications·Chan CaoYi-Tao Long
Oct 31, 2018·Biosensors·Marta Maria Pereira da Silva Neves, Daniel Martín-Yerga
Jan 21, 2021·Life·Ana CrnkovićGregor Anderluh
Feb 3, 2021·Biochimica Et Biophysica Acta. Biomembranes·Motahareh Ghahari LarimiLiviu Movileanu
Aug 21, 2019·Biophysical Journal·Bach PhamMin Chen
Sep 17, 2013·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Marco BacciFabio Cecconi
May 11, 2021·Biochimica Et Biophysica Acta. Biomembranes·David P HoogerheideSergey M Bezrukov
Jul 3, 2021·Biosensors·Bo LinHongju Mao
Mar 30, 2019·ACS Nano·Motahareh Ghahari LarimiLiviu Movileanu

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