# Tuning and Tagging the O-GlcNAc Modification

> **NIH NIH U01** · JOHNS HOPKINS UNIVERSITY · 2020 · $445,400

## Abstract

Summary
 Thousands of intracellular proteins are dynamically modified by monosaccharides of O-linked N-
acetylglucosamine (O-GlcNAc). The cycling of O-GlcNAc is regulated by the concerted actions of enzymes
encoded by just two genes: the O-GlcNAc transferase (OGT) and the O-GlcNAcase (OGA) that add and remove
O-GlcNAc, respectively (1-4). Dysregulation of O-GlcNAc cycling, or levels of the nucleotide sugar used by OGT
(UDP-GlcNAc), exacerbates the pathophysiology of a host of diseases including type II diabetes, cancer,
neurodegeneration, heart failure, hypertension and aging (5-7). While technological and methodological
innovations have improved our ability to detect, modulate, and site-map O-GlcNAc (8-13), many of these
techniques have not been adopted by the broader scientific community thus inhibiting a mechanistic
understanding of the roles that O-GlcNAc plays in potentiating disease. Often, the aforementioned approaches
require specialized equipment and reagents, lack the specificity required to study a modification that is cycled
by just two enzymes, or lead to significant off-target effects (10, 14-20). Thus, the goal of the studies proposed
herein is to generate facile, inexpensive, chemical genetic tools that probe the roles of O-GlcNAc in vitro and in
vivo. Specifically, we will address the following aims:
 Specific Aim #1: Enabling tunable, reversible chemical-genetic regulation of O-GlcNAcylation in an
organelle-specific manner. These tools will combine destabilization domain (21-23) and nanotrap (24)
technology to enable researchers to modulate the expression of OGT and OGA, as well as key enzymes within
the hexosamine biosynthetic pathway, in a spatial, temporal, and dose-dependent manner. These tools will be
applied to understanding the role of O-GlcNAc in prostate cancer.
 Specific Aim 2: Enabling tissue- and organelle-specific tagging of O-GlcNAc-modified proteins. This
tool will harness the power of proximity Biotin ligation (25, 26) to enable tagging and enrichment of O-
GlcNAcylated proteins in a cell-specific or organelle-specific manner.
 Together, the tools described in this proposal will overcome current experimental limitations associated with
studying O-GlcNAc and facilitate studies focused on determining the role of O-GlcNAc at a mechanistic level in
a broad range of models.

## Key facts

- **NIH application ID:** 9948600
- **Project number:** 5U01CA230978-03
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Natasha Elizabeth Zachara
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $445,400
- **Award type:** 5
- **Project period:** 2018-08-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9948600

## Citation

> US National Institutes of Health, RePORTER application 9948600, Tuning and Tagging the O-GlcNAc Modification (5U01CA230978-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9948600. Licensed CC0.

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