Detergent-free extraction of a functional low-expressing GPCR from a human cell line

Publication date: Available online 13 December 2019Source: Biochimica et Biophysica Acta (BBA) - BiomembranesAuthor(s): Juan Francisco Bada Juarez, Juan C. Muñoz-García, Rosana Inácio dos Reis, Alistair Henry, David McMillan, Marco Kriek, Martyn Wood, Catherine Vandenplas, Zara Sands, Luis Castro, Richard Taylor, Anthony WattsAbstractDopamine receptors (DRs) are class A G-Protein Coupled Receptors (GPCRs) prevalent in the central nervous system (CNS). These receptors mediate physiological functions ranging from voluntary movement and reward recognition to hormonal regulation and hypertension. Drugs targeting dopaminergic neurotransmission have been employed to treat several neurological and psychiatric disorders, including Parkinson's disease, schizophrenia, Huntington's disease, attention deficit hyperactivity disorder (ADHD), and Tourette's syndrome. In vivo, incorporation of GPCRs into lipid membranes is known to be key to their biological function and, by inference, maintenance of their tertiary structure. A further significant challenge in the structural and biochemical characterization of human DRs is their low levels of expression in mammalian cells. Thus, the purification and enrichment of DRs whilst retaining their structural integrity and function is highly desirable for biophysical studies. A promising new approach is the use of styrene–maleic acid (SMA) copolymer to solubilize GPCRs directly in their native environment, to produce polymer-assembled Lipodisqs ...
Source: Biochimica et Biophysica Acta (BBA) Biomembranes - Category: Biochemistry Source Type: research