# Solid-state NMR studies of the dynamic interactions of the influenza A M2 membrane protein with water, antiviral drugs, and the M1 protein

> **NIH NIH F31** · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · 2020 · $38,065

## Abstract

Abstract
 Seasonal influenza viruses infect 5-10% of adults and 20-30% of children each year, yet only two classes
of antiviral drugs are available so far, one targeting the influenza A M2 (AM2) protein and the other targeting
neuraminidase (NA). AM2 forms a proton channel in the virus envelope that is crucial for the virus lifecycle;
blockage of the AM2 pore curbs virus infection. Unfortunately, in recent years 99% of flu strains have become
resistant to the FDA-approved AM2 channel blockers, amantadine and rimantadine. Recently, new antiviral
drugs effective against amantadine-resistant AM2 proteins have been reported, but their mechanisms of action
have remained elusive. Water plays an important role in proton conduction through the AM2 pore, but it is not
understood how the new AM2 inhibitors affect water dynamics in the channel. Elucidating channel water
dynamics should provide insights into the mechanism of action of anti-influenza drugs. I propose to develop and
employ solid-state nuclear magnetic resonance (SSNMR) spectroscopy to investigate the dynamic interactions
of AM2 with water, with drugs, and with another virus protein, matrix protein 1 (AM1). Specifically, I will develop
SSNMR techniques that correlate the dynamically sensitive 2H quadrupolar coupling with 13C chemical shifts to
provide site-resolved information about small-molecule dynamics and protein dynamics. In Aim 1, I will measure
the effects of antiviral drugs on channel hydration and on water dynamics in the AM2 pore. Both 1H-13C and 2H-
13C correlation experiments will be conducted to detect the dynamics of protonated and deuterated channel
water, respectively, giving complementary information. The results will provide a detailed molecular
understanding of the mechanism of action of the novel drugs and facilitate future design of better antiviral
compounds. In Aim 2, I will measure the conformational dynamics of AM2 as influenced by drugs, to address
how the transmembrane (TM) domain dynamics affect antiviral drug activity and whether the protein dynamics
is related to channel-water dynamics. These studies will shed light on the yet unkown binding site of recently
developed antiviral drugs. 2H, 13C-labeled proteins will be used in this study. Secondly, I will characterize AM2
interaction with AM1. Biochemical evidence suggests that this interaction is important for virus assembly and
budding, thus elucidating the structural basis for this interaction may help the design of alternative antiviral drugs.
I will measure changes in the AM2 cytoplasmic domain’s dynamics and chemical shifts upon binding to AM1, to
understand whether binding immobilizes AM2 and changes protein structure. I have obtained extensive
preliminary data that demonstrate the feasibility of the 2H-13C correlation techniques and have successfully
expressed and purified 13C,2H,15N-labeled proteins. It will also provide first-time information about the importance
of molecular motion on the function of an i...

## Key facts

- **NIH application ID:** 9996472
- **Project number:** 5F31AI133989-03
- **Recipient organization:** MASSACHUSETTS INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Martin David Gelenter
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $38,065
- **Award type:** 5
- **Project period:** 2018-09-01 → 2021-05-14

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9996472

## Citation

> US National Institutes of Health, RePORTER application 9996472, Solid-state NMR studies of the dynamic interactions of the influenza A M2 membrane protein with water, antiviral drugs, and the M1 protein (5F31AI133989-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9996472. Licensed CC0.

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