Year: 2013
Type: article
Abstract: Protein nanopore-based sensing elements represent a pressing need in molecular biomedical diagnosis (Bayley & Cremer, 2001). However, the integration of protein nanopores with other nanofluidic devices is a challenging task. This is especially true, if we consider that isolated single proteins are in general fragile and unstable under harsh conditions of detection. Here, I will present a strategy for improving the stability of the open-state current of a redesigned nanopore using ferric hydroxamate uptake component A (FhuA), a beta-barrel membrane protein channel of E. coli (Mohammad et al., 2012). The primary function of FhuA is to facilitate the energy-driven, high-affinity Fe3+ uptake complexed by the siderophore ferrichrome (Paweleket al., 2006). The key ingredient of this strategy was the coupling of direct genetic engineering of FhuA with a fast-dilution refolding approach to obtain an unusually stable protein nanopore under a broad range of experimentation. These advantageous characteristics were recently demonstrated by examining proteolytic activity of an enzyme at a highly acidic pH, a condition at which majority of beta-barrel protein nanopores are normally gated or unfolded. Future membrane protein design work will not only reveal a better understanding of the processes employed in membrane protein folding and stability, but will also serve as a platform for the integration of robust protein components into devices (Astier, Bayley, & Howorka, 2005).
Author: L. Movileanu
Institution: Syracuse University
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: bronze
OpenAlex ID: W2012220710