Biomedical Engineering Reference
In-Depth Information
Chapter 9
Nanopore Recordings to Quantify
Activity-Related Properties of Proteins
Erik C. Yusko, Yazan N. Billeh, Jerry Yang, and Michael Mayer
Abstract Electrical current recordings through electrolyte-filled nanopores (so called
resistive pulse-sensing experiments) are attracting increasing attention for identifying
and characterizing biomolecules. The majority of the work employing this method
so far has focused on detection of oligonucleotides, polymers, and viruses. Most
recently nanopores have been used to detect single proteins. This chapter reviews
the very first attempts to use nanopores for characterizing properties of proteins that
relate to their activity. The emphasis lies on those studies that provided quantitative
information on activity-related properties of proteins, such as protein conformation,
ligand binding, and enzyme activity. Nanopore-based studies have tremendous poten-
tial for investigating the function of proteins because the technique is capable of
interrogating individual proteins at high-throughput without requiring labeling.
Keywords Protein • Virus • Nucleotide • Polymer • Protein conformation • Protein
volume • Protein charge • Electrophoretic mobility • Drift velocity • Isoelectric
point • Ligand affinity • Stoichiometry • Association constant • Dissociation
constant • Kinetics • On-rate • Off-rate • Binding isotherm • Drug-protein interac-
tion • Bovine serum albumin • Ovalbumin • Avidin • Streptavidin • Antibody
• Immunoglobulin G • Immunoglobulin E • Antibody fab fragments • Staphylococ-
cal enterotoxin B •
b
-Lactoglobulin • E.
coli
maltose binding protein • Lectin
• Carbonic anhydrase II • Sulfonamide • RNA aptamer • Fibrinogen • Poly (ethylene
glycol) • Biotin • Ricin • Enzyme activity • Protease • Phospholipase • Membrane-
active enzyme • Catalytic rate constant • Forward rate constant • Michaelis
constant • Phospholipase C • Phospholipase D • Anthrax lethal factor • Trypsin
• Alkaline phosphatase • Amyloid-
b
• Phosphatidylcholine • Phosphatidylinositol
•
-hemolysin • Gramicidin • Alamethicin • Resistive pulse • Current blockage
• Translocation time • Sojourn time • Biosensing • BLM • Planar lipid bilayer
a
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