# Real Time (RT) In-cell NMR technology to study protein interactions in live cells

> **NIH NIH R01** · STATE UNIVERSITY OF NEW YORK AT ALBANY · 2020 · $307,911

## Abstract

Real Time (RT) In-cell NMR technology to study protein interactions in live cells
Alexander Shekhtman1
1Department of Chemistry, University at Albany, State University of New York, Albany, NY 12222
ABSTRACT: Since intracellular changes induced by stimuli can take hours to fully manifest into
detectable signals, one of the overarching goals of modern biology is to understand temporal protein
structure-function relationships at atomic resolution within the complexity of a live cell. In-cell solution
NMR spectroscopy is an important step towards this goal but is limited by the long data acquisition times
and the static nature of in-cell NMR experiments that provide snapshots rather than continuous
monitoring of time dependent changes in protein structure. In addition, weak quinary interactions between
the protein of interest and intracellular components, particularly RNAs, which are omnipresent in live
cells, result in a dramatic increase in the apparent in-cell molecular weight and render all but a few
proteins invisible by standard in-cell NMR approaches. There are currently no structural biology tools to
characterize time dependent protein interactions in live cells at atomic resolution even though these
interactions affect protein physicochemical properties, protein-protein, protein-ligand, and protein-drug
binding. We showed that the combination of protein deuteration and NMR experiments using optimized
transverse relaxation allowed us to obtain in-cell NMR spectra of previously invisible proteins. The goal
of this project is to develop real time (RT) in-cell NMR technology to characterize protein interactions in
situ over a long (more than a day) period of time at atomic resolution inside live prokaryotic and
eukaryotic cells. We will apply this technology to study how exogenous and endogenous challenges to
cells result in specific temporal changes in protein structure. We will build the infrastructure to make this
new technology available to the scientific community. We expect that the technology will be critical to
bridge the gap between in vitro and in-cell protein biochemistry, which is an absolute requirement to
understand cell biology and to develop effective therapeutics against protein targets.

## Key facts

- **NIH application ID:** 9912182
- **Project number:** 5R01GM085006-08
- **Recipient organization:** STATE UNIVERSITY OF NEW YORK AT ALBANY
- **Principal Investigator:** ALEXANDER SHEKHTMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $307,911
- **Award type:** 5
- **Project period:** 2010-07-01 → 2022-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9912182

## Citation

> US National Institutes of Health, RePORTER application 9912182, Real Time (RT) In-cell NMR technology to study protein interactions in live cells (5R01GM085006-08). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9912182. Licensed CC0.

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