자료유형  

 학위논문

Control Number  

0017162403

International Standard Book Number  

9798383567425

Dewey Decimal Classification Number  

574

Main Entry-Personal Name  

Critcher, Megan L.

Publication, Distribution, etc. (Imprint  

[S.l.] : The Scripps Research Institute., 2024

Publication, Distribution, etc. (Imprint  

Ann Arbor : ProQuest Dissertations & Theses, 2024

Physical Description  

185 p.

General Note  

Source: Dissertations Abstracts International, Volume: 86-01, Section: B.

General Note  

Advisor: Huang, Mia L.

Dissertation Note  

Thesis (Ph.D.)--The Scripps Research Institute, 2024.

Summary, Etc.  

요약To expand the functional repertoire of their proteome, higher organisms have evolved protein post-translational modifications (PTMs), including glycosylation. Proteoglycans (PGs) are unique glycan-modified proteins demarcated by the presence of covalently attached poly-sulfated, O-linked glycosaminoglycan (GAG) chains. PGs serve crucial roles in (1) signal transduction, (2) host-pathogen interactions, and (3) interactions within the extracellular matrix (ECM). Although PGs are now well regarded as key facilitators of cell biology, their structure-function relationships remain difficult to study and largely unknown due to their complex structure.This complexity arises from their multivariate nature; each PG is defined by a unique core protein sequence but can embody a collection of structural variants due to differences in the (1) number of GAGs, (2) identity, and sulfation of GAGs, and (3) membrane localization. Through these various structural elements, PGs can mediate and fine-tune a range of biological processes. Traditional chemical or genetic approaches fail to control the full complexity of PGs, inhibited by limitations of chemical synthesis and off-target effects. I postulated that a chemical biology approach combining methods from both would be uniquely suited to address the vast chemical complexity of these conjugates and consider both protein and glycan components equally.Here, I present a novel method incorporating of synthetic carbohydrate derivatives, unnatural amino acid incorporation and click chemistry to create de novo PGs with controlled glycosylation and cell surface localization. With these glycoconjugates, I demonstrate the ability to comprehensively study PG binding interactions in vitro and in cellulo with fine-scale control over PG structure. I additionally utilized these constructs to characterize the binding preferences of a monoclonal antibody targeting the PG syndecan-1 (SDC1). Lastly, I postulated that the collection of PGs present on the cell surface could be elucidated through proximity labeling with a GAG-binding protein (GAGBP). By appending the ascorbate peroxidase APEX2 to fibroblast growth factor 2 (FGF2), I mapped its interactome in two biologically relevant cell lines to identify cell type-specific PGs. Through this project, I hope to identify the functionally relevant PGs that are required for FGF2 signal transduction through in cellulo knockdown experiments.

Subject Added Entry-Topical Term  

Cellular biology.

Subject Added Entry-Topical Term  

Biology.

Subject Added Entry-Topical Term  

Biochemistry.

Index Term-Uncontrolled  

Glycobiology

Index Term-Uncontrolled  

Glycosaminoglycans

Index Term-Uncontrolled  

Heparan sulfate

Index Term-Uncontrolled  

Proteoglycans

Index Term-Uncontrolled  

Host-pathogen interactions

Added Entry-Corporate Name  

The Scripps Research Institute Chemical Biology

Host Item Entry  

Dissertations Abstracts International. 86-01B.

Electronic Location and Access  

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Control Number  

joongbu:658460