# Sparse NMR Labeling Approach to Glycoprotein Structure and Function

> **NIH NIH R01** · UNIVERSITY OF GEORGIA · 2022 · $302,000

## Abstract

SUMMARY
 Glycoproteins represent a class of mammalian proteins that presents challenges for structural and
functional characterization, particularly if the native glycosylation, which affects structure, stability and
interaction with other molecules, is to be preserved. The best route to proteins with native glycosylation is
expression in mammalian cell cultures. Unfortunately, for X-ray crystallography, this produces proteins with
heterogenous glycosylation, often preventing formation of suitable crystals. For traditional nuclear magnetic
resonance (NMR) methods, this forces use of expensive substrates for isotopic labeling and prohibits the
perdeuteration often required to maintain resolution for larger proteins. The investigators involved in this
proposal have worked together to develop an efficient mammalian cell expression system that produces
proteins sparsely labeled using a restricted set of less expensive isotope enriched amino acids and maintains
resolution without the aid of perdeuteration. This has been accompanied by the development of resonance
assignment programs and data analysis protocols that allow structural and functional characterization from
basic, high sensitivity, two-(and three-)dimensional NMR experiments. This project is designed to turn those
developments into an integrated protocol that can be adopted by an expanded community of users. It centers
on the refinement of a software package that accomplishes NMR resonance assignment of sparsely labeled
proteins. It will be bolstered by extensive validation of program output, introduction of new data types and data
analysis methods, and extension of program capabilities to the refinement of computer-generated models for
protein structure. The potential impact will be a new route to structure and function studies of a class of
proteins intimately involved with human physiology and disease.

## Key facts

- **NIH application ID:** 10388355
- **Project number:** 5R01GM134335-04
- **Recipient organization:** UNIVERSITY OF GEORGIA
- **Principal Investigator:** JAMES H. PRESTEGARD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $302,000
- **Award type:** 5
- **Project period:** 2019-07-01 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10388355

## Citation

> US National Institutes of Health, RePORTER application 10388355, Sparse NMR Labeling Approach to Glycoprotein Structure and Function (5R01GM134335-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10388355. Licensed CC0.

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