Crystallizing Membrane Proteins for Structure Determination using Lipidic Mesophases

Review the theoretical background and rationale for using lipidic mesophases in membrane protein crystallization trials. This foundation supports understanding of subsequent preparation and handling steps.

Assemble all necessary equipment, materials, and reagents for the crystallization setup, including crystallization plates, syringes, lipids, precipitants, and protein samples.

Set up and organize the crystallization plate with appropriate wells and protective measures to prevent contamination and evaporation during sample dispensing.

Mix the membrane protein with lipid in the correct stoichiometry and concentration, ensuring homogeneous dispersion in the lipidic mesophase before loading into syringes.

Transfer the prepared lipid-protein mixture into the dispensing syringe using precise techniques to avoid air bubbles and ensure accurate dosing of crystallization samples.

Carefully dispense the lipid-protein mixture onto the prepared crystallization plates following the established protocol to create conditions suitable for crystal formation.

Document initial crystal appearance and growth patterns following dispensing, establishing baseline conditions for subsequent structure determination experiments.