A fast, simple and robust protocol for growing crystals in the lipidic cubic phase

Area of Science:

• Structural Biology
• Biochemistry
• Crystallography

Background:

• Membrane protein crystallization is crucial for understanding their function.
• The lipidic cubic mesophase (in meso method) is a challenging yet effective technique for membrane protein crystallization.
• Existing protocols can be complex and time-consuming, posing a barrier to entry for researchers.

Purpose of the Study:

• To describe a simple, inexpensive, and rapid protocol for in meso membrane protein crystallization.
• To facilitate researcher familiarity and comfort with the lipidic cubic mesophase.
• To enable the crystallization of valuable membrane and soluble proteins.

Main Methods:

• Development of a straightforward protocol for crystal production within the lipidic cubic mesophase.
• Utilizing glass sandwich plates for crystal preparation.
• Testing the protocol with lysozyme as a model protein.

Main Results:

• Crystals formed rapidly, within 15-30 minutes at 293 K.
• The protocol proved effective for lysozyme crystallization, demonstrating its utility.
• Prepared samples are suitable for practicing crystal harvesting, cooling, and diffraction data collection.

Conclusions:

• This robust protocol significantly simplifies the in meso method for membrane protein crystallization.
• It lowers the barrier to entry for researchers, enabling broader application of the technique.
• The protocol supports subsequent steps in the crystallization workflow, including data collection.