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Solubilization and reconstitution of the mu-opioid receptor expressed in human neuronal SH-SY5Y and CHO cells
- Source :
- Peptides. 55
- Publication Year :
- 2014
-
Abstract
- The zwitterionic detergent CHAPS was used to solubilize the human mu-opioid receptor (hMOR) from SH-SY5Y neuroblastoma cells and recombinant hMOR-CHO (CHO-T7-hMOR) and hMOR-SH-SY5Y (SH-SY5Y-T7-hMOR) cell membranes. Agonist stimulation and G-protein activation by the mu-selective opioid agonist DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin) were recovered after removing of CHAPS after polyethylene glycol (PEG) precipitation. Binding assays show that hMOR solubilized and reconstituted this way was functional and able to interact with both agonist peptides and with G-protein. The effective solubilization and reconstitution of hMOR from mammalian cells, without truncation and extensive modification, represent an essential step toward the purification of a receptor bearing important post-translational modifications.
- Subjects :
- Agonist
SH-SY5Y
Physiology
medicine.drug_class
Narcotic Antagonists
Detergents
Receptors, Opioid, mu
Diprenorphine
CHO Cells
Ligands
Biochemistry
Protein Refolding
Polyethylene Glycols
Cellular and Molecular Neuroscience
chemistry.chemical_compound
Endocrinology
Cricetulus
Chaps
Cell Line, Tumor
medicine
Animals
Humans
Receptor
G protein-coupled receptor
Chemistry
Chinese hamster ovary cell
Cholic Acids
Enkephalin, Ala(2)-MePhe(4)-Gly(5)
DAMGO
Solubility
Guanosine 5'-O-(3-Thiotriphosphate)
μ-opioid receptor
Protein Binding
Subjects
Details
- ISSN :
- 18735169
- Volume :
- 55
- Database :
- OpenAIRE
- Journal :
- Peptides
- Accession number :
- edsair.doi.dedup.....331e3da08282fdfd2fa748b367fb6deb