Isoprenoid-chained lipid EROCOC₁₇₊₄: a new matrix for membrane protein crystallization and a crystal delivery medium in serial femtosecond crystallography.

ABSTRACT: In meso crystallization of membrane proteins relies on the use of lipids capable of forming a lipidic cubic phase (LCP). However, almost all previous crystallization trials have used monoacylglycerols, with 1-(cis-9-octadecanoyl)-rac-glycerol (MO) being the most widely used lipid. We now report that EROCOC₁₇₊₄ mixed with 10% (w/w) cholesterol (Fig. 1) serves as a new matrix for crystallization and a crystal delivery medium in the serial femtosecond crystallography of Adenosine A_2A receptor (A_2AR). The structures of EROCOC₁₇₊₄-matrix grown A_2AR crystals were determined at 2.0 Å resolution by serial synchrotron rotation crystallography at a cryogenic temperature, and at 1.8 Å by LCP-serial femtosecond crystallography, using an X-ray free-electron laser at 4 and 20 °C sample temperatures, and are comparable to the structure of the MO-matrix grown A_2AR crystal (PDB ID: 4EIY). Moreover, X-ray scattering measurements indicated that the EROCOC₁₇₊₄/water system did not form the crystalline L_C phase at least down to -?20 °C, in marked contrast to the equilibrium MO/water system, which transforms into the crystalline L_C phase below about 17 °C. As the L_C phase formation within the LCP-matrix causes difficulties in protein crystallography experiments in meso, this feature of EROCOC₁₇₊₄ will expand the utility of the in meso method.

SUBMITTER: Ihara K 

PROVIDER: S-EPMC7652841 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature