Does the lipid environment impact the open-state conductance of an engineered β-barrel protein nanopore?

Noriko Tomita, Mohammad M. Mohammad, David J. Niedzwiecki, Makoto Ohta, Liviu Movileanu

Research output: Contribution to journalArticle

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Abstract

Using rational membrane protein design, we were recently able to obtain a β-barrel protein nanopore that was robust under an unusually broad range of experimental circumstances. This protein nanopore was based upon the native scaffold of the bacterial ferric hydroxamate uptake component A (FhuA) of Escherichia coli. In this work, we expanded the examinations of the open-state current of this engineered protein nanopore, also called FhuA ΔC/Δ4L, employing an array of lipid bilayer systems that contained charged and uncharged as well as conical and cylindrical lipids. Remarkably, systematical single-channel analysis of FhuA ΔC/Δ4L indicated that most of its biophysical features, such as the unitary conductance and the stability of the open-state current, were not altered under the conditions tested in this work. However, electrical recordings at high transmembrane potentials revealed that the presence of conical phospholipids within the bilayer catalyzes the first, stepwise current transition of the FhuA ΔC/Δ4L protein nanopore to a lower-conductance open state. This study reinforces the stability of the open-state current of the engineered FhuA ΔC/Δ4L protein nanopore under various experimental conditions, paving the way for further critical developments in biosensing and molecular biomedical diagnosis.

Original languageEnglish (US)
Pages (from-to)1057-1065
Number of pages9
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1828
Issue number3
DOIs
StatePublished - Mar 2013

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Nanopores
Lipids
Proteins
Motor Skills
Bibliography of Medicine
Lipid Bilayers
Membrane Potentials
Phospholipids
Membrane Proteins
Escherichia coli
Frangula
Scaffolds

Keywords

  • Channel closure
  • Dimensionless shape factor
  • E. coli FhuA
  • Lipid effect
  • Planar lipid bilayer
  • Single-channel recording

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Biophysics

Cite this

Does the lipid environment impact the open-state conductance of an engineered β-barrel protein nanopore? / Tomita, Noriko; Mohammad, Mohammad M.; Niedzwiecki, David J.; Ohta, Makoto; Movileanu, Liviu.

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1828, No. 3, 03.2013, p. 1057-1065.

Research output: Contribution to journalArticle

Tomita, Noriko; Mohammad, Mohammad M.; Niedzwiecki, David J.; Ohta, Makoto; Movileanu, Liviu / Does the lipid environment impact the open-state conductance of an engineered β-barrel protein nanopore?

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1828, No. 3, 03.2013, p. 1057-1065.

Research output: Contribution to journalArticle

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