Off-campus UMass Amherst users: To download campus access dissertations, please use the following link to log into our proxy server with your UMass Amherst user name and password.
Non-UMass Amherst users: Please talk to your librarian about requesting this dissertation through interlibrary loan.
Dissertations that have an embargo placed on them will not be available to anyone until the embargo expires.
Author ORCID Identifier
N/A
AccessType
Open Access Dissertation
Document Type
dissertation
Degree Name
Doctor of Philosophy (PhD)
Degree Program
Chemistry
Year Degree Awarded
2019
Month Degree Awarded
February
First Advisor
Min Chen
Subject Categories
Biochemistry | Biological Engineering | Biomaterials | Molecular Biology | Molecular, Cellular, and Tissue Engineering | Other Biochemistry, Biophysics, and Structural Biology
Abstract
The use of pore-forming proteins (PFPs) in nanopore sensing has been fruitful largely due to their nanoscale size and the ease with which protein nanopores can be manipulated and consistently reproduced at a large scale. Nanopore sensing relies heavily on a steady ionic current afforded by rigid nanopores, as the change in current is indicative of analyte detection, revealing characteristics of the analyte such as its relative size, concentration, and charge, as well as the nanopore:analyte interaction. Rigid PFPs have been used in applications such as DNA sequencing, kinetic studies, analyte discrimination, and protein conformation dynamics at the single-molecule level. The work in this dissertation details a different approach to nanopore sensing using a non-rigid PFP, the monomeric outer membrane protein G (OmpG), which exhibits intrinsic current fluctuations called gating. We exploit the gating of OmpG for the effective detection and discrimination of protein homologues and isoforms, showing that OmpG gating is particularly useful in selectively detecting targets and probing nanopore:analyte interactions. Further, we demonstrate the ability of OmpG to retain its sensing capabilities in complex mixtures of serum and human cell lysate.
DOI
https://doi.org/10.7275/13382695
Recommended Citation
Yang, Bib, "Building the Outer Membrane Protein G (OmpG) Nanopore Library: From the Discrimination of Biotin-Binding Proteins in Serum to Resolving Human Carbonic Anhydrase From Human Red Blood Cells" (2019). Doctoral Dissertations. 1513.
https://doi.org/10.7275/13382695
https://scholarworks.umass.edu/dissertations_2/1513
Included in
Biochemistry Commons, Biological Engineering Commons, Biomaterials Commons, Molecular Biology Commons, Molecular, Cellular, and Tissue Engineering Commons, Other Biochemistry, Biophysics, and Structural Biology Commons