title: Uptake of the butyrate receptors, GPR41 and GPR43, in lipidic bicontinuous cubic phases suitable for in meso crystallization.
abstract: The butyrate G-protein coupled receptors (GPCRs), GPR41 and GPR43, have been implicated in colorectal cancer and leptin production. To date their function has not been elucidated as low levels of protein expression and difficulties in producing diffraction quality crystals have hindered their structural determination. In meso crystallization, which uses an artificial lipid membrane matrix to facilitate crystal growth, is becoming an increasingly successful crystallization technique, particularly for GPCRs. We report herein the lipid membrane matrix structural characterization for GPR41 and GPR43 within two lipid self-assembly systems (monoolein and phytantriol) commonly used for in meso crystallization and comment on their suitability for crystallizing these GPCRs. Synchrotron small angle X-ray scattering (SAXS) studies were used to determine the initial phase and uptake of these receptors within the lipid matrix and investigate the role of cholesterol in this process. The self-assembled lipid nanostructure was retained in the presence of GPR43 for both lipids but was destabilized for GPR41 in the phytantriol lipid system. The structural changes to the lipid matrix upon protein incorporation were greater for cholesterol-doped systems, potentially indicative of increased receptor uptake.
authors.affiliation: Monash University, Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, VIC 3052, Australia.
authors.forename: Yi-Lynn
authors.initials: YL
authors.lastname: Liang
authors.affiliation: RMIT, School of Applied Sciences, College of Science, Engineering and Health, GPO Box 2476, Melbourne, VIC 3001, Australia; CSIRO Materials Science and Engineering (CMSE), Private Bag 10, Clayton South MDC, VIC 3169, Australia.
authors.forename: Charlotte E
authors.initials: CE
authors.lastname: Conn
authors.affiliation: RMIT, School of Applied Sciences, College of Science, Engineering and Health, GPO Box 2476, Melbourne, VIC 3001, Australia; CSIRO Materials Science and Engineering (CMSE), Private Bag 10, Clayton South MDC, VIC 3169, Australia.
authors.forename: Calum J
authors.initials: CJ
authors.lastname: Drummond
authors.affiliation: CSIRO Materials Science and Engineering (CMSE), 385 Royal Parade, Parkville, VIC 3052, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, La Trobe University, Bundoora, VIC 3086, Australia. Electronic address: c.darmanin@latrobe.edu.au.
authors.forename: Connie
authors.initials: C
authors.lastname: Darmanin
mesh_terms: D003460:Crystallization; D005233:Fatty Alcohols / Q000737:chemistry; D005989:Glycerides / Q000737:chemistry; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008958:Models, Molecular; D044367:Phase Transition; D011956:Receptors, Cell Surface / Q000737:chemistry*; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D005233:Fatty Alcohols; D005989:Glycerides; D008055:Lipids; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C473103:FFAR2 protein, human; C106262:FFAR3 protein, human; C508873:3,7,11,15-tetramethyl-1,2,3-hexadecanetriol; C005953:monoolein
keywords: Bicontinuous cubic phase; Butyrate receptor; GPCR; GPR41; GPR43; In meso crystallization; Monoolein; Phytantriol