Literature DB >> 34085261

Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor.

Alexei A Yeliseev1.   

Abstract

G protein-coupled receptors (GPCR) are integral membrane proteins that regulate multiple cellular processes. To obtain insights into structural properties of GPCR and mechanism of activity, these proteins should be isolated in significant (milligram) quantities, in a pure, homogenous, and stable form. Here we describe the expression and purification of type II human cannabinoid receptor CB2, a class A GPCR, in two different types of expression hosts: in Escherichia coli and in mammalian suspension cell culture Expi293. Our method allows preparation of milligram quantities of the purified receptors suitable for a wide array of downstream applications including high-resolution structural studies and functional assays.

Entities:  

Keywords:  CB2 receptor; Membrane protein; Recombinant expression; Thermostability

Year:  2021        PMID: 34085261     DOI: 10.1007/978-1-0716-1221-7_4

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  18 in total

1.  Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused.

Authors:  R B Kapust; D S Waugh
Journal:  Protein Sci       Date:  1999-08       Impact factor: 6.725

2.  Use of dual affinity tags for expression and purification of functional peripheral cannabinoid receptor.

Authors:  Alexei Yeliseev; Lioudmila Zoubak; Klaus Gawrisch
Journal:  Protein Expr Purif       Date:  2006-12-12       Impact factor: 1.650

3.  Development of the Twin-Strep-tag® and its application for purification of recombinant proteins from cell culture supernatants.

Authors:  Thomas G M Schmidt; Lilia Batz; Lidia Bonet; Uwe Carl; Gerd Holzapfel; Klaus Kiem; Kamila Matulewicz; Dennis Niermeier; Isabel Schuchardt; Kristian Stanar
Journal:  Protein Expr Purif       Date:  2013-09-06       Impact factor: 1.650

4.  Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency.

Authors:  R B Kapust; J Tözsér; J D Fox; D E Anderson; S Cherry; T D Copeland; D S Waugh
Journal:  Protein Eng       Date:  2001-12

Review 5.  Immobilized-metal affinity chromatography (IMAC): a review.

Authors:  Helena Block; Barbara Maertens; Anne Spriestersbach; Nicole Brinker; Jan Kubicek; Roland Fabis; Jörg Labahn; Frank Schäfer
Journal:  Methods Enzymol       Date:  2009       Impact factor: 1.600

6.  Mutagenesis of a flexible loop in streptavidin leads to higher affinity for the Strep-tag II peptide and improved performance in recombinant protein purification.

Authors:  S Voss; A Skerra
Journal:  Protein Eng       Date:  1997-08

7.  Probing thermostability of detergent-solubilized CB2 receptor by parallel G protein-activation and ligand-binding assays.

Authors:  Ryan L Beckner; Lioudmila Zoubak; Kirk G Hines; Klaus Gawrisch; Alexei A Yeliseev
Journal:  J Biol Chem       Date:  2019-11-27       Impact factor: 5.157

8.  Characterization of Small Isotropic Bicelles with Various Compositions.

Authors:  K S Mineev; K D Nadezhdin; S A Goncharuk; A S Arseniev
Journal:  Langmuir       Date:  2016-06-22       Impact factor: 3.882

9.  A case of invasive papillary breast carcinoma: Fierce façade with favorable prognosis.

Authors:  D Vani; S Geetanjali; Giri M Punja; M Bharathi
Journal:  J Cancer Res Ther       Date:  2015 Oct-Dec       Impact factor: 1.805

10.  Use of tandem affinity chromatography for purification of cannabinoid receptor CB₂.

Authors:  Silvia C Locatelli-Hoops; Alexei A Yeliseev
Journal:  Methods Mol Biol       Date:  2014
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  1 in total

1.  Production of Eukaryotic Glycoproteins for Structural and Functional Studies Using Expi293F Cells.

Authors:  Jessica A Greven; Tom J Brett
Journal:  Curr Protoc       Date:  2022-08
  1 in total

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