# Atomic Resolution Characterization of Kindlin-2 Binding to Phosphatidylinositol Phosphatases in Lipid Bilayers by Solid-State NMR

> **NIH NIH R01** · RUTGERS, THE STATE UNIV OF N.J. · 2022 · $324,804

## Abstract

Project Summary:
Kindlin-2 (K2) is a peripheral membrane protein which regulates integrin, a key protein that mediates cellular
adhesion in humans. The goal of this project is to increase our structural understanding of K2 and the lipid
membranes to which it binds, to provide key insights into the regulatory function of K2. The interactions of K2
with cellular membranes are difficult to study because membranes are highly dynamic and lack long-range order.
As a result, our ability to study membranes and membrane proteins have been limited despite their potential to
be impactful in a wide array of systems. In this proposal we will focus on the signaling lipids phosphatidylinositol
phosphates (PIPs) which control the activity of K2. This lipid family is important for human health due to the role
of PIPs as regulators of numerous cell processes including nutrient sensing and growth. The misregulation of
PIP-dependent pathways is implicated in numerous rare and common diseases, including cancer and diabetes.
Mechanistic details of PIP activation and regulation of protein binding partners, like K2, are essential to
understanding and combating these diseases. Solid-state NMR is a unique tool for the atomic resolution
characterization of the structure and dynamics of lipids and determining the factors that govern lipid-protein
interactions. Over the five-year timeline of this proposal I plan to develop NMR experiments for directly
investigating K2-lipid interfaces with high sensitivity and specificity. The focus of this study will be the conserved
PIP binding domains of K2: a ubiquitin-like F0 domain and a pleckstrin homology (PH) domain. The presence of
naturally NMR-active and chemically distinct phosphate groups in the PIPs will serve as a handle to directly
probe the binding domain-lipid interface. Characterization of specific interactions between lipid head groups and
proteins will be the foundation upon which we examine the structural basis of PIP recognition and the mechanism
of K2 activation by PIPs. Beyond studies of the protein-lipid interface, we seek to understand how membrane
binding affects the interactions of K2 with partner proteins and itself. Our experiments will take advantage of all
the latest improvements in NMR hardware, particularly very-fast magic angle spinning. My group and I are highly
experienced in developing novel experiments to assign the chemical shifts and solve structures of protein
complexes, membrane proteins, and protein fibrils. This provides us with the tools that are required to tackle the
structural aspects of lipid-protein and protein-protein interfaces. We will use an integrative approach in which
state-of-the-art computational techniques will guide our experimental work. All-atom molecular dynamics will be
used to provide hypotheses that may be tested with NMR and to provide atomic resolution interpretations of
experimental data. This work is vital for assisting in the design of therapeutics and diagnostics tha...

## Key facts

- **NIH application ID:** 10492704
- **Project number:** 5R01GM139905-02
- **Recipient organization:** RUTGERS, THE STATE UNIV OF N.J.
- **Principal Investigator:** Andrew Nieuwkoop
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $324,804
- **Award type:** 5
- **Project period:** 2021-09-30 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10492704

## Citation

> US National Institutes of Health, RePORTER application 10492704, Atomic Resolution Characterization of Kindlin-2 Binding to Phosphatidylinositol Phosphatases in Lipid Bilayers by Solid-State NMR (5R01GM139905-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10492704. Licensed CC0.

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