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Author ORCID Identifier

https://orcid.org/0000-0002-5896-1600

AccessType

Campus-Only Access for Five (5) Years

Document Type

dissertation

Degree Name

Doctor of Philosophy (PhD)

Degree Program

Molecular and Cellular Biology

Year Degree Awarded

2021

Month Degree Awarded

September

First Advisor

Scott C. Garman

Second Advisor

Daniel N. Hebert

Third Advisor

Margaret M. Stratton

Fourth Advisor

Eric R. Strieter

Subject Categories

Biochemistry | Biophysics | Molecular Biology | Structural Biology

Abstract

Lysosomal storage diseases affect about 1 in 5000 live births but most of them remain without treatment options. Fabry disease is commonly treated with enzyme replacement therapy, but kidney pathology remains problematic. We have engineered a smaller form of the treatment enzyme that should increase the concentration of protein that is able to reach a key part of the kidney filtration barrier, the podocytes. Our enzyme retains the ability to cleave Gb3, retains activity better than wild type in conditions relevant to enzyme replacement therapy and remains 99% identical to the wild type protein sequence. For another glycosidase deficiency, fucosidosis, we have established an expression system for the glycosylated form of alpha-L-fucosidase and demonstrate the potential of deoxyfuconojirimycin (DFJ) and fucose as pharmacological chaperones. Both chaperones were able to increase the lysosomal colocalization of the disease mutant S155F and both greatly stabilize the wild type enzyme at both acidic and neutral pH. These studies provide initial insights into two important treatment options for fucosidosis and warrant deeper investigations of both in cellular and animal models of the disease.

DOI

https://doi.org/10.7275/24369892

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Available for download on Thursday, September 01, 2022

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