Engineering a rigid protein tunnel for biomolecular detection

Mohammad M. Mohammad, Raghuvaran Iyer, Khalil R. Howard, Mark P. McPike, Philip N. Borer, Liviu Movileanu

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)9521-9531
Number of pages11
JournalJournal of the American Chemical Society
Volume134
Issue number22
DOIs
StatePublished - Jun 6 2012

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Proteins
Nanopores
Motor Skills
Membrane Proteins
Communication Disorders
Tunnels
Membranes
Nucleotide Aptamers
Genetic Engineering
Nanostructures
Peptide Hydrolases
Salts
Ions
Technology
Acyclic Acids
Bibliography of Medicine
Dental Pulp Calcification
Beta-Globulins
Scaffolds
Kinetics

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Mohammad, M. M., Iyer, R., Howard, K. R., McPike, M. P., Borer, P. N., & Movileanu, L. (2012). Engineering a rigid protein tunnel for biomolecular detection. Journal of the American Chemical Society, 134(22), 9521-9531. DOI: 10.1021/ja3043646

Engineering a rigid protein tunnel for biomolecular detection. / Mohammad, Mohammad M.; Iyer, Raghuvaran; Howard, Khalil R.; McPike, Mark P.; Borer, Philip N.; Movileanu, Liviu.

In: Journal of the American Chemical Society, Vol. 134, No. 22, 06.06.2012, p. 9521-9531.

Research output: Contribution to journalArticle

Mohammad, MM, Iyer, R, Howard, KR, McPike, MP, Borer, PN & Movileanu, L 2012, 'Engineering a rigid protein tunnel for biomolecular detection' Journal of the American Chemical Society, vol 134, no. 22, pp. 9521-9531. DOI: 10.1021/ja3043646
Mohammad MM, Iyer R, Howard KR, McPike MP, Borer PN, Movileanu L. Engineering a rigid protein tunnel for biomolecular detection. Journal of the American Chemical Society. 2012 Jun 6;134(22):9521-9531. Available from, DOI: 10.1021/ja3043646

Mohammad, Mohammad M.; Iyer, Raghuvaran; Howard, Khalil R.; McPike, Mark P.; Borer, Philip N.; Movileanu, Liviu / Engineering a rigid protein tunnel for biomolecular detection.

In: Journal of the American Chemical Society, Vol. 134, No. 22, 06.06.2012, p. 9521-9531.

Research output: Contribution to journalArticle

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