ArticleJournal of molecular biology2011
Engineering an ultra-thermostable β(1)-adrenoceptor.
Article in Journal of molecular biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 53 citations in OpenAlex.
- TSH receptor autoantibodies - a personal experience.Thyroid research · 2026Review
- Autoantibodies to the TSH Receptor-from discovery to understanding the mechanisms of action and to new therapeutics.Endocrine journal · 2025Review
- Filling of a water-free void explains the allosteric regulation of the βNature chemistry · 2022Article
- Stabilization and structure determination of integral membrane proteins by termini restraining.Nature protocols · 2022Review
- Efficient production of a functional G protein-coupled receptor in E. coli for structural studies.Journal of biomolecular NMR · 2021Article
- GPCR Activation States Induced by Nanobodies and Mini-G Proteins Compared by NMR Spectroscopy.Molecules (Basel, Switzerland) · 2020Article
- IMPROvER: the Integral Membrane Protein Stability Selector.Scientific reports · 2020Article
- Insights into the Basal Activity and Activation Mechanism of the β1 Adrenergic Receptor Using Native Mass Spectrometry.Journal of the American Society for Mass Spectrometry · 2019Article
- Towards high throughput GPCR crystallography: In Meso soaking of Adenosine AScientific reports · 2018Article
- Engineering a minimal G protein to facilitate crystallisation of G protein-coupled receptors in their active conformation.Protein engineering, design & selection : PEDS · 2016Article
- How Do Short Chain Nonionic Detergents Destabilize G-Protein-Coupled Receptors?Journal of the American Chemical Society · 2016Article
- A mutagenesis and screening strategy to generate optimally thermostabilized membrane proteins for structural studies.Nature protocols · 2016Article
- Generating Recombinant Antibodies to Membrane Proteins through Phage Display.Antibodies (Basel, Switzerland) · 2016Review
- Stabilization of a prokaryotic LAT transporter by random mutagenesis.The Journal of general physiology · 2016Article
- How Can Mutations Thermostabilize G-Protein-Coupled Receptors?Trends in pharmacological sciences · 2016Review
- Pharmacological Analysis and Structure Determination of 7-Methylcyanopindolol-Bound β1-Adrenergic Receptor.Molecular pharmacology · 2015Article
- An economic approach to efficient isotope labeling in insect cells using homemade 15N-, 13C- and 2H-labeled yeast extracts.Journal of biomolecular NMR · 2015Article
- Identification of destabilizing and stabilizing mutations of Ste2p, a G protein-coupled receptor in Saccharomyces cerevisiae.Biochemistry · 2015Article
- Stabilization of G protein-coupled receptors by point mutations.Frontiers in pharmacology · 2015Review
- Large-scale production and protein engineering of G protein-coupled receptors for structural studies.Frontiers in pharmacology · 2015Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Conformational thermostabilisation of G-protein-coupled receptors is a successful strategy for their structure determination. The thermostable mutants tolerate short-chain detergents, such as octylglucoside and nonylglucoside, which are ideal for crystallography, and in addition, the receptors are preferentially in a single conformational state. The first thermostabilised receptor to have its structure determined was the β(1)-adrenoceptor mutant β(1)AR-m23 bound to the antagonist cyanopindolol, and recently, additional structures have been determined with agonist bound. Here, we describe further stabilisation of β(1)AR-m23 by the addition of three thermostabilising mutations (I129V, D322K, and Y343L) to make a mutant receptor that is 31 °C more thermostable than the wild-type receptor in dodecylmaltoside and is 13 °C more thermostable than β(1)AR-m23 in nonylglucoside. Although a number of thermostabilisation methods were tried, including rational design of disulfide bonds and engineered zinc bridges, the two most successful strategies to improve the thermostability of β(1)AR-m23 were an engineered salt bridge and leucine scanning mutagenesis. The three additional thermostabilising mutations did not significantly affect the pharmacological properties of β(1)AR-m23, but the new mutant receptor was significantly more stable in short-chain detergents such as heptylthioglucoside and denaturing detergents such as SDS.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.