Open AccessJournal Article
Electrostatic Interactions between OmpG Nanopore and Analyte Protein Surface Can Distinguish between Glycosylated Isoforms
Monifa A. Fahie,Min Chen +1 more
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TLDR
It is shown that OmpG nanopore equipped with a biotin ligand can distinguish glycosylated and deglycosylation isoforms of avidin by their differences in surface charge.Abstract:
The flexible loops decorating the entrance of OmpG nanopore move dynamically during ionic current recording. The gating caused by these flexible loops changes when a target protein is bound. The gating is characterized by parameters including frequency, duration, and open-pore current, and these features combine to reveal the identity of a specific analyte protein. Here, we show that OmpG nanopore equipped with a biotin ligand can distinguish glycosylated and deglycosylated isoforms of avidin by their differences in surface charge. Our studies demonstrate that the direct interaction between the nanopore and analyte surface, induced by the electrostatic attraction between the two molecules, is essential for protein isoform detection. Our technique is remarkably sensitive to the analyte surface, which may provide a useful tool for glycoprotein profiling.read more
Citations
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Functionalizing nanoparticles with cancer-targeting antibodies: A comparison of strategies.
TL;DR: This review summarizes adsorption, covalent conjugation and biotin-avidin interaction, while discussing the advantages, limitations and relevant therapeutic approaches currently under investigation.
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Nanopore-Based Single-Biomolecule Interfaces: From Information to Knowledge.
TL;DR: The design of the nanopore-based single biomolecule interface is outlined, which provides rich stochastic information regarding each biomolecules, and the concept of "sin-gle-molecule ionic spectrum" is discussed, which may allow mapping of noncovalent interactions at an atomic level in the future.
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Biological Nanopores: Confined Spaces for Electrochemical Single-Molecule Analysis.
Chan Cao,Yi-Tao Long +1 more
TL;DR: The application of biological nanopores are introduced to investigate the conformations of peptides affected by charge, length, and dipole moment and to study disease-related proteins' structures and aggregation transitions influenced by an inhibitor, a promoter, or an applied voltage.
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Label-Free Detection of Post-translational Modifications with a Nanopore
TL;DR: It is shown that phosphorylated and glycosylated peptides can be clearly differentiated from nonmodified peptides by differences in the relative current blockade and dwell time in nanopore translocations, an important step for the single-molecule, label-free identification of proteoforms, which have tremendous potential for disease diagnosis and cell biology.
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Selective Detection of Protein Homologues in Serum Using an OmpG Nanopore
TL;DR: The results demonstrate the feasibility of directly profiling proteins in real-world samples with minimal or no sample pretreatment and show that two antibiotin antibodies each induced a distinct gating pattern that allowed them to be readily detected and simultaneously discriminated by a single OmpG nanopore in the presence of fetal bovine serum.
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