Abstract
With the advent of the recent determination of high-resolution crystal structures of bovine rhodopsin and human β2 adrenergic receptor (β2AR), there are still many structure-function relationships to be learned from other G protein- coupled receptors (GPCRs). Many of the pharmaceutically interesting GPCRs cannot be modeled because of their amino acid sequence divergence from bovine rhodopsin and β2AR. Structure determination of GPCRs can provide new avenues for engineering drugs with greater potency and higher specificity. Several obstacles need to be overcome before membrane protein structural biology becomes routine: over-expression, solubilization, and purification of milligram quantities of active and stable GPCRs. Coordinated iterative efforts are required to generate any significant GPCR overexpression. To formulate guidelines for GPCR purification efforts, we review published conditions for solubilization and purification using detergents and additives. A discussion of sample preparation of GPCRs in detergent phase, bicelles, nanodiscs, or low-density lipoproteins is presented in the context of potential structural biology applications. In addition, a review of the solubilization and purification of successfully crystallized bovine rhodopsin and β2AR highlights tools that can be used for other GPCRs.
Combinatorial Chemistry & High Throughput Screening
Title: Over-Expression, Solubilization, and Purification of G Protein-Coupled Receptors for Structural Biology
Volume: 11 Issue: 6
Author(s): Mark L. Chiu, Cindy Tsang, Nelson Grihalde and Maria P. MacWilliams
Affiliation:
Abstract: With the advent of the recent determination of high-resolution crystal structures of bovine rhodopsin and human β2 adrenergic receptor (β2AR), there are still many structure-function relationships to be learned from other G protein- coupled receptors (GPCRs). Many of the pharmaceutically interesting GPCRs cannot be modeled because of their amino acid sequence divergence from bovine rhodopsin and β2AR. Structure determination of GPCRs can provide new avenues for engineering drugs with greater potency and higher specificity. Several obstacles need to be overcome before membrane protein structural biology becomes routine: over-expression, solubilization, and purification of milligram quantities of active and stable GPCRs. Coordinated iterative efforts are required to generate any significant GPCR overexpression. To formulate guidelines for GPCR purification efforts, we review published conditions for solubilization and purification using detergents and additives. A discussion of sample preparation of GPCRs in detergent phase, bicelles, nanodiscs, or low-density lipoproteins is presented in the context of potential structural biology applications. In addition, a review of the solubilization and purification of successfully crystallized bovine rhodopsin and β2AR highlights tools that can be used for other GPCRs.
Export Options
About this article
Cite this article as:
Chiu L. Mark, Tsang Cindy, Grihalde Nelson and MacWilliams P. Maria, Over-Expression, Solubilization, and Purification of G Protein-Coupled Receptors for Structural Biology, Combinatorial Chemistry & High Throughput Screening 2008; 11 (6) . https://dx.doi.org/10.2174/138620708784911456
DOI https://dx.doi.org/10.2174/138620708784911456 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Radiofrequency Radiation and Human Triiodothronine Hormone: Immunoenzymometric Assay
Recent Patents on Biomarkers Dendrimer Based Anti-Infective and Anti-Inflammatory Drugs
Recent Patents on Anti-Infective Drug Discovery Inhibitors of Chronically Active Ras: Potential for Treatment of Human Malignancies
Recent Patents on Anti-Cancer Drug Discovery Fundamentals of Prion Diseases and Their Involvement in the Loss of Function of Cellular Prion Protein
Protein & Peptide Letters Manipulation of Glycolysis in Malignant Tumors: Fantasy or Therapy?
Current Medicinal Chemistry An Expanding Appreciation of the Role Chemokine Receptors Play in Cancer Progression
Current Pharmaceutical Design Editorial (Thematic Issue: Focal Adhesion Kinase and Cross-Linked Signaling In Cancer)
Anti-Cancer Agents in Medicinal Chemistry miRNA-497 Enhances the Sensitivity of Colorectal Cancer Cells to Neoadjuvant Chemotherapeutic Drug
Current Protein & Peptide Science The Targets of Curcumin
Current Drug Targets Immunotherapy with Tumor Vaccines for the Treatment of Malignant Gliomas
Current Drug Discovery Technologies Nano-Delivery in Pediatric Tumors: Looking Back, Moving Forward
Anti-Cancer Agents in Medicinal Chemistry Disintegrins from Snake Venoms and their Applications in Cancer Research and Therapy
Current Protein & Peptide Science CDK9 a Potential Target for Drug Development
Medicinal Chemistry Prolyl Oligopeptidase: A Rising Star on the Stage of Neuroinflammation Research
CNS & Neurological Disorders - Drug Targets The Role of 3D Pharmacophore Mapping Based Virtual Screening for Identification of Novel Anticancer Agents: An Overview
Current Topics in Medicinal Chemistry Epigenetic Effects of Cadmium in Cancer: Focus on Melanoma
Current Genomics Sodium Selenite Decreased HDAC Activity, Cell Proliferation and Induced Apoptosis in Three Human Glioblastoma Cells
Anti-Cancer Agents in Medicinal Chemistry The Clinical Impact of Quantitative Cell-free DNA, KRAS, and BRAF Mutations on Response to Anti-EGFR Treatment in Patients with Metastatic Colorectal Cancer
Current Pharmaceutical Design HER2 in the Era of Molecular Medicine: A Review
Current Cancer Therapy Reviews EDITORIAL [Hot topic: New Therapeutic Advances and Perspectives in Tumour Angiogenesis (Guest Editor: Eddy Pasquier)]
Current Cancer Drug Targets