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Updated: May 17, 2026

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

Published on: March 10, 2023

High-throughput methods for electron crystallography.

David L Stokes¹, Iban Ubarretxena-Belandia, Tamir Gonen

¹Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)

|November 8, 2012

Summary

This summary is machine-generated.

This study introduces two high-throughput methods for screening membrane protein crystallization for electron crystallography. These techniques enhance the study of membrane proteins, crucial for drug development and understanding cell physiology.

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High-Throughput Protein Crystallization via Microdialysis

High-Throughput Protein Crystallization via Microdialysis

Published on: March 3, 2023

Area of Science:

Structural Biology
Biophysics
Biochemistry

Background:

Membrane proteins are vital for cell function and drug targets.
Their complex nature hinders structural studies compared to soluble proteins.
Electron crystallography offers native lipid environments for membrane protein structure determination.

Purpose of the Study:

To develop high-throughput screening methods for membrane protein crystallization in electron crystallography.
To overcome limitations in current crystallization screening for membrane proteins.
To facilitate the structural determination of membrane proteins for drug discovery.

Main Methods:

Developed a 96-well dialysis format using liquid-handling robots for detergent removal and bilayer reconstitution.
Implemented a cyclodextrin titration method with a specialized pipetting robot and light scattering for monitoring reconstitution.
Utilized liquid-handling robots for grid preparation and automated imaging in electron microscopy.

Main Results:

Achieved high-throughput screening for membrane protein crystallization trials.
Demonstrated efficient detergent removal and bilayer reconstitution using novel methods.
Enabled systematic screening and monitoring of crystallization processes.

Conclusions:

The developed methods significantly increase throughput for electron crystallography screening of membrane proteins.
These advancements facilitate structural studies of membrane proteins, aiding in drug development.
Automated processes improve efficiency and reproducibility in membrane protein structural biology.