# Purchase of helium recovery system for NMR spectrometers

> **NIH NIH R01** · TEXAS A&M AGRILIFE RESEARCH · 2020 · $25,042

## Abstract

Project Summary
The pandemic 1918 influenza A virus (IAV), also known as the Spanish flu, caused the worst influenza pandemic
in human history. Nonstructural protein 1 (NS1) is a multifunctional virulence factor associated with the
suppression of anti-viral immune responses and thereby has been identified as one of the molecular
determinants of high pathogenicity of the 1918 IAV. NS1 of the 1918 IAV (1918 NS1) contains a proline-rich
motif (PRM) that mediates binding with host CrkII with high affinity and selectivity. The 1918 NS1:CrkII interaction
plays critical roles in the suppression of host anti-viral immune responses and the enhancement of viral
replication. Moreover, NS1s of many avian/swine IAVs contain the CrkII-binding PRM. Given the zoonotic
potential of IAVs, there is a critical need to determine the molecular mechanisms by which the interaction of 1918
NS1 and cellular CrkII is regulated. The long-term goal of our research program is to elucidate the molecular
mechanisms underlying virus-host protein interactions. Our objectives in this proposal are to determine the
structural mechanisms of the 1918 NS1:CrkII interaction, and to determine the molecular and cellular
mechanisms whereby the 1918 NS1:CrkII complex induces PI3K activation, resulting in enhanced viral
replication. Our central hypothesis is that the 1918 NS1:CrkII complex is structurally dynamic, which is
functionally important for the interaction with the p85 regulatory subunit of PI3K. To test this hypothesis, we will
determine the structure of the 1918 NS1:CrkII complex and elucidate how the complex interacts with p85 to
activate the PI3K signaling pathway. Our rationale for these studies is that the mechanistic understanding of the
interactions of 1918 NS1 with CrkII and p85 would help identify previously undiscovered target sites to develop
for potential inhibitors against the 1918 NS1. Through a synergistic approach combining small-angle X-ray
scattering, NMR spectroscopy, molecular dynamics simulation, and cell-based assays, we will pursue the
following specific aims. Aim 1. To determine the structural mechanism of the 1918 NS1:CrkII interaction using a
battery of biophysical experiments. Hijacking and relocation of CrkII into the nucleus is a distinctive feature of
the 1918 pandemic IAV NS1. To understand this process, we will reveal structural and energetic mechanisms
by which the affinity and lifetime (1/koff) of the 1918 NS1:CrkII complex are modulated. Aim 2. To determine the
molecular mechanism underlying NS1-induced PI3K activation. The 1918 NS1:CrkII interaction markedly
enhances NS1-induced PI3K activation; however, its molecular mechanism is unknown. We will seek to
comprehensively determine the molecular mechanisms by which the 1918 NS1:CrkII complex interacts with the
p85 subunit of PI3K, reveal its functional role in PI3K activation, and identify hotspot NS1 residues that interact
with both CrkII and p85. This study is expected to have a positive im...

## Key facts

- **NIH application ID:** 10135522
- **Project number:** 3R01GM127723-02S2
- **Recipient organization:** TEXAS A&M AGRILIFE RESEARCH
- **Principal Investigator:** Jae Hyun Cho
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $25,042
- **Award type:** 3
- **Project period:** 2018-09-20 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10135522

## Citation

> US National Institutes of Health, RePORTER application 10135522, Purchase of helium recovery system for NMR spectrometers (3R01GM127723-02S2). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10135522. Licensed CC0.

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