# Technology Development 2:  MAS NMR and dynamic nuclear polarization for HIV-1 structural biology

> **NIH NIH P50** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2020 · $228,350

## Abstract

Abstract for Technology Development Program 2 - MAS NMR
It has become clear that dynamics within protein molecules comprising HIV-1 assemblies play critical roles in
regulating viral infectivity, including uncoating and maturation. Current progress in the field is hampered by the
paucity of structural-biological methods that yield atomic-level information into structure and dynamics
simultaneously, particularly when large-amplitude conformational rearrangements take place. Magic angle
spinning (MAS) NMR is uniquely positioned to yield such information and, when integrated with molecular
dynamics (MD) simulations, provides dynamic information inaccessible by any other means. While MAS NMR is a
very powerful technique, it currently suffers from three major drawbacks: i) inherent low sensitivity, resulting in
long measurement times; ii) high spectral congestion for large assemblies due to numerous overlapping signals; and
iii) extensive and time-consuming data analysis for large systems (resonance assignments and structure
calculation), hampering the widespread use of the method.
We propose to develop a new methodological framework for atomic-level structural, dynamic, and mechanistic
characterization of HIV-1 protein assemblies by MAS NMR, which will overcome the main roadblocks, limited
sensitivity and resolution, as well as long data analysis time. To accomplish this goal, we will integrate high
magnetic fields (17.6–28.2 T) with ultrafast MAS frequencies, proton detection, streamlined data acquisition,
processing, and analysis. We will also employ dynamic nuclear polarization (DNP)-based experiments for analysis
of low-concentration species as well as for investigations of the conformational space accessible to the dynamically
disordered states. We will develop new experiments suitable for studies of large assemblies of HIV-1 proteins and
their complexes with host proteins and small-molecule interactors, in a fraction of time and with a small fraction of
material that is required for conventional experiments.
The dramatic sensitivity and/or resolution enhancements foreseen to become attainable through the combination
of proposed MAS NMR and DNP methods will greatly expand the range of systems amenable to in-depth
characterization. The streamlined data analysis through the integration of experiment and computation will
dramatically improve the throughput of MAS NMR and make the technique accessible to a very wide cohort of
researchers. The technologies developed for the analysis of HIV-1 assemblies through the proposed studies will be
broadly applicable to other biological assemblies. Finally, we envision that the proposed methodological framework
will pave the way for the atomic-resolution structural and dynamics characterization of viral proteins in the context
of intact viruses.

## Key facts

- **NIH application ID:** 9977963
- **Project number:** 5P50AI150481-14
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Tatyana Polenova
- **Activity code:** P50 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $228,350
- **Award type:** 5
- **Project period:** 2007-08-27 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9977963

## Citation

> US National Institutes of Health, RePORTER application 9977963, Technology Development 2:  MAS NMR and dynamic nuclear polarization for HIV-1 structural biology (5P50AI150481-14). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/9977963. Licensed CC0.

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