# Dynamics of membrane proteins unraveled by time-resolved serial crystallography

> **NIH NIH R01** · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · 2023 · $383,684

## Abstract

ABSTRACT
Membrane proteins are of extreme importance for human health and 30% of the human proteome consist of
membrane proteins, which are key players in important cellular processes controlling and mediating the
interaction between cells, regulate transport in and out of the cells and are also the major players in bioenergy
conversion including respiration; 40% of all current drugs being targeted to membrane proteins. However
knowledge of their structure and function at the atomic level is sparse, as the structures for less 800 unique
membrane proteins has been determined to date. Furthermore, most of these structures show only a static
picture of the membrane protein, while their function in the cell is highly dynamic. This proposal aims to
develop novel methods to determine “molecular movies” of membrane proteins “at work,” and specifically, to
determine the dynamics of the catalytic cycle of the cytochrome oxidase and to study the conformational
dynamics of the beta-adrenergic G-protein coupled receptor. The proposal aims to develop and use a
combination of methods that focus on time-resolved femtosecond (fs) X-ray crystallography with XFELs. Fs
crystallography, pioneered by our team in the previous funding cycles, has revolutionized X-ray
crystallography. It overcomes radiation damage, enabling structure analysis of biological macromolecules at
room temperature under physiological conditions based on the new serial approach for structure determination,
where tens of thousands of X-ray diffraction snapshots are collected from a stream of fully hydrated
nano/microcrystals of proteins, interacting with fs X-ray pulses from a Free Electron Laser. New developments
in nanocrystal growth and characterization, together with development of new injector technology and data
evaluation methods, have progressed the new method of SFX at a very fast pace based on results from this
project, which has led to 70 publications, 29 of them in high impact journals as well as patents (accepted and
provisional applications filed). This R01 Renewal is based on the success of the previous work but explores
new areas by shifting the major focus from the proof-of-concept to their improvement and applicationtowards
molecular movies of the functional dynamics of two important membrane proteins: cytochrome c oxidase and
the beta-adrenergic receptor. In Aim 1 we will study the dynamics of the catalytic cycle of cytochrome c
oxidase, a large-multi-protein membrane protein complex and the key enzyme in respiration, catalyzing the 4
electron 4 proton reduction of oxygen to 2 water molecules, whose detailed mechanisms is still a hot topic of
debate. Aim 2 is focused on the dynamics of the beta-adrenergic receptor and its complexes with arrestin and
G-protein. In this Aim, we will combine new SFX method developments (including making the GPCR triggered
by light to allow for very fast dynamics to be detected) with the time resolved cryo-EM studies of the GPCR
complexes that wi...

## Key facts

- **NIH application ID:** 10657320
- **Project number:** 5R01GM095583-12
- **Recipient organization:** ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
- **Principal Investigator:** PETRA FROMME
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $383,684
- **Award type:** 5
- **Project period:** 2010-09-30 → 2025-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10657320

## Citation

> US National Institutes of Health, RePORTER application 10657320, Dynamics of membrane proteins unraveled by time-resolved serial crystallography (5R01GM095583-12). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10657320. Licensed CC0.

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