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Insights into the mechanism of high lipid–detergent crystallization of membrane proteins

DOI: 10.1107/S1600576721010669 DOI Help

Authors: Sofia Trampari (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine) , Caroline Neumann (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine) , Samuel J. Hjorth-Jensen (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine) , Azadeh Shahsavar (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine) , Esben M. Quistgaard (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine) , Poul Nissen (Aarhus University; DANDRITE, Nordic EMBL Partnership for Molecular Medicine)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Applied Crystallography , VOL 54

State: Published (Approved)
Published: December 2021
Diamond Proposal Number(s): 23316

Abstract: Obtaining well diffracting crystals of membrane proteins is often challenging, but chances can be improved by crystallizing them in lipidic conditions that mimic their natural membrane environments. One approach is the high lipid–detergent (HiLiDe) method, which works by mixing the target protein with high concentrations of lipid and detergent prior to crystallization. Although this approach is convenient and flexible, understanding the effects of systematically varying lipid/detergent ratios and a characterization of the lipid phases that form during crystallization would be useful. Here, a HiLiDe phase diagram is reported for the model membrane protein MhsT, which tracks the precipitation and crystallization zones as a function of lipid and detergent concentrations, and is augmented with data on crystal sizes and diffraction properties. Additionally, the crystallization of SERCA1a solubilized directly with native lipids is characterized as a function of detergent concentration. Finally, HiLiDe crystallization drops are analysed with transmission electron microscopy, which among other features reveals liposomes, stacked lamellae that may represent crystal precursors, and mature crystals with clearly discernible packing arrangements. The results emphasize the significance of optimizing lipid/detergent ratios over broad ranges and provide insights into the mechanism of HiLiDe crystallization.

Journal Keywords: X-ray crystallography; membrane proteins; lipid phases; high lipid–detergent (HiLiDe) method; crystallization phase diagrams; electron microscopy

Subject Areas: Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography

Other Facilities: P14 beamline at PETRA III

Added On: 26/11/2021 10:10

Discipline Tags:

Life Sciences & Biotech Structural biology

Technical Tags:

Diffraction Macromolecular Crystallography (MX)