# Breaking barriers in CryoEM through computational protein design

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA-IRVINE · 2022 · $372,060

## Abstract

ABSTRACT
Using single-particle cryogenic electron microscopy (cryoEM) we can study proteins and protein complexes to
atomic resolution and elucidate dynamics and distinct functional states in native or near-native environments.
But this is virtually impossible to do with small proteins. My lab is interested in understanding protein function by
observing their structures at high-resolution in native states. We leverage expertise in cryoEM and computational
protein design to study proteins, both soluble and membrane, and their complexes that play major roles in human
diseases. Membrane proteins are cellular gatekeepers and one of the most important class of membrane
proteins are the G protein-coupled receptors (GPCRs), which act as signal conduits through ligand-induced
binding on outside cells to the recruitment of binding complexes inside cells to relay signals. As they are involved
in most cellular processes they are of considerable interest in drug development.
 While cryoEM has gained momentum in structural biology, it has a fundamental limitation that proteins
smaller than 40 kDa cannot be studied effectively because the signal in the images is too low. This means that
most proteins in the human genome cannot be studied by cryoEM. Using a designed approach, I was recently
able to solve the high-resolution structure of a 17kDa protein by cryoEM, almost 3 times smaller than current
cryoEM size limits. I succeeded because we used computational design to attach the 17 kDa protein to a scaffold
which helped imaging by increasing the mass of the particle and the higher symmetry afforded better
reconstruction. This proof-of-principal experiment demonstrates the powerful combination of using computational
design for cryoEM. While exciting, this scaffold approach is still at its infancy and further design and development
are needed to realize its full potential. New scaffolds capable of displaying important membrane proteins like
GPCRs will be developed, tested and optimized. These new nanomaterials will be specifically tailored to address
cryoEM needs. With this approach we will investigate membrane protein structure, describe functional dynamics
in near native environments and facilitate rapid structure-guided drug design to help against devastating
diseases.

## Key facts

- **NIH application ID:** 10470905
- **Project number:** 5R35GM142797-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA-IRVINE
- **Principal Investigator:** Shane Gonen
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $372,060
- **Award type:** 5
- **Project period:** 2021-09-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10470905

## Citation

> US National Institutes of Health, RePORTER application 10470905, Breaking barriers in CryoEM through computational protein design (5R35GM142797-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10470905. Licensed CC0.

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