DOI
https://doi.org/10.25772/95PD-HG83
Author ORCID Identifier
https://orcid.org/0000-0002-6094-7373
Defense Date
2022
Document Type
Dissertation
Degree Name
Doctor of Philosophy
Department
Pharmaceutical Sciences
First Advisor
Umesh Desai
Abstract
Glycosaminoglycans (GAGs) are linear, carbohydrate polymers comprising significant structural diversity in chain length and pattern of sulfation. GAGs are able to interact with hundreds of currently known proteins and modulate the activity of many. In some cases, GAGs are able to bind proteins with high affinity and selectivity primarily due to specific patterns of sulfation that are recognized by the proteins. Microarray-based technology has proven to be an effective tool for studying GAGs and their interactions with various biomolecular entities. Here, we utilize a heparan sulfate (HS) microarray to screen for relevant structure–function relationships in the binding of HS to SARS-CoV-2 spike glycoprotein (SgP). From this, sulfation patterns including sulfation at the 3-O-position on glucosamine residues was determined to be significant. The HS microarray was also screened against the receptor tyrosine kinase (RTK) IGF-1R and its cognate ligand IGF-1 to elucidate the system’s dependencies on chain length and sulfation pattern for HS molecules. Finally, we constructed two structurally distinct GAG-based microarrays using separate in-house developed methodologies. These microarrays were screened against a number of proteins including SARS-CoV-2 SgP and its variant forms as well as RTKs including IGF-1R, and relevant structure–function relationships were posited for these systems. This work includes extensive application of GAG microarray technology as a vehicle to better understanding relevant GAG–protein interactions in biology by way of identifying the structure-function relationships that allow for said molecular events.
Rights
© John E. Chittum
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
12-12-2022
Included in
Biophysics Commons, Biotechnology Commons, Cancer Biology Commons, Immunology of Infectious Disease Commons, Molecular Biology Commons