6LPJ

Isoprenoid-chained lipid EROCOC 17+4 : a new matrix for membrane protein crystallization and a crystal delivery medium in serial femtosecond crystallography.

(2020) Sci Rep 10: 19305-19305

• PubMed: 33168855 Search on PubMedSearch on PubMed Central
• DOI: https://doi.org/10.1038/s41598-020-76277-x
• Primary Citation of Related Structures:  
  6LPJ, 6LPK, 6LPL


• PubMed 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 EROCOC ₁₇₊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 _2A R). The structures of EROCOC ₁₇₊₄ -matrix grown A _2A R 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 _2A R crystal (PDB ID: 4EIY). Moreover, X-ray scattering measurements indicated that the EROCOC ₁₇₊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 EROCOC ₁₇₊₄ will expand the utility of the in meso method.

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Organizational Affiliation: 

RIKEN Center for Biosystems Dynamics Research, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.