title: Real-time shape approximation and fingerprinting of single proteins using a nanopore.
abstract: Established methods for characterizing proteins typically require physical or chemical modification steps or cannot be used to examine individual molecules in solution. Ionic current measurements through electrolyte-filled nanopores can characterize single native proteins in an aqueous environment, but currently offer only limited capabilities. Here we show that the zeptolitre sensing volume of bilayer-coated solid-state nanopores can be used to determine the approximate shape, volume, charge, rotational diffusion coefficient and dipole moment of individual proteins. To do this, we developed a theory for the quantitative understanding of modulations in ionic current that arise from the rotational dynamics of single proteins as they move through the electric field inside the nanopore. The approach allows us to measure the five parameters simultaneously, and we show that they can be used to identify, characterize and quantify proteins and protein complexes with potential implications for structural biology, proteomics, biomarker detection and routine protein analysis.
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Erik C
authors.identifier: 0000-0003-3559-8375
authors.initials: EC
authors.lastname: Yusko
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Brandon R
authors.initials: BR
authors.lastname: Bruhn
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.|Adolphe Merkle Institute, University of Fribourg, Chemin des Verdiers 4, CH-1700 Fribourg, Switzerland.
authors.forename: Olivia M
authors.initials: OM
authors.lastname: Eggenberger
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.|Adolphe Merkle Institute, University of Fribourg, Chemin des Verdiers 4, CH-1700 Fribourg, Switzerland.
authors.forename: Jared
authors.initials: J
authors.lastname: Houghtaling
authors.affiliation: Department of Physics, University of Arkansas, Fayetteville, Arkansas 72701, USA.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Rollings
authors.affiliation: Department of Physics, University of Arkansas, Fayetteville, Arkansas 72701, USA.
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Walsh
authors.affiliation: Department of Physics, University of Arkansas, Fayetteville, Arkansas 72701, USA.
authors.forename: Santoshi
authors.initials: S
authors.lastname: Nandivada
authors.affiliation: Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut 06269, USA.
authors.forename: Mariya
authors.initials: M
authors.lastname: Pindrus
authors.affiliation: Department of Biomedical Engineering and Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston Salem, North Carolina 27157, USA.
authors.forename: Adam R
authors.initials: AR
authors.lastname: Hall
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.|Center for Computational Medicine and Biology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: David
authors.initials: D
authors.lastname: Sept
authors.affiliation: Department of Physics, University of Arkansas, Fayetteville, Arkansas 72701, USA.
authors.forename: Jiali
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut 06269, USA.
authors.forename: Devendra S
authors.initials: DS
authors.lastname: Kalonia
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.|Adolphe Merkle Institute, University of Fribourg, Chemin des Verdiers 4, CH-1700 Fribourg, Switzerland.|Biophysics Program, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Mayer
mesh_terms: D008051:Lipid Bilayers / Q000737:chemistry*; D008956:Models, Chemical; D046912:Multiprotein Complexes / Q000737:chemistry*; D058608:Nanopores
chemical_list: D008051:Lipid Bilayers; D046912:Multiprotein Complexes