Structure basis of lipophilic ligand binding of membrane protein involving in lipid mediator biosynthesis

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H04343
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Ago Hideo 国立研究開発法人理化学研究所, 放射光科学総合研究センター, 専任研究員 (70360477)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: Protein crystallography / membrane protein / lipid mediator

Outline of Final Research Achievements

Structure basis of lipophilic ligand recognition of human membrane proteins involving in lipid mediator biosynthesis was the subject of this study. The membrane proteins utilize hydrophilic GSH and lipophilic arachidonic acid metabolites. In comparison to GSH whose binding architecture was elucidated in detail by X-ray crystallography, limited information about lipophilic ligand was available. This study used two methods to get the lipophilic ligand / protein complex, one was the soaking method on the crystals grown in surfo and the other was the co-crystallization in meso. In addition to making lipophilic ligand / protein complex, a method spreading thinly a highly viscus vehicle in which a large number of micro-crystals were dispersed was developed. Crystal overlap along X-ray beam causes the double hit which makes the diffraction image unavailable. The thin spreading reduces the crystal overlap; thus, resulting in the efficient data collection from micro crystals.

Free Research Field

Synchrotron structural biology