Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies
Membrane proteins such as G protein-coupled receptors (GPCRs) exert fundamental biological functions and are involved in a multitude of physiological responses, making these receptors ideal drug targets. Drug discovery programs targeting GPCRs have been greatly facilitated by the emergence of high-r...
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doaj-32bcdb1efb6c4f54a379fb8c496ae1a02021-03-09T00:04:07ZengMDPI AGMolecules1420-30492021-03-01261465146510.3390/molecules26051465Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical StudiesYann Waltenspühl0Janosch Ehrenmann1Christoph Klenk2Andreas Plückthun3Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zürich, SwitzerlandDepartment of Biochemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zürich, SwitzerlandDepartment of Biochemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zürich, SwitzerlandDepartment of Biochemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zürich, SwitzerlandMembrane proteins such as G protein-coupled receptors (GPCRs) exert fundamental biological functions and are involved in a multitude of physiological responses, making these receptors ideal drug targets. Drug discovery programs targeting GPCRs have been greatly facilitated by the emergence of high-resolution structures and the resulting opportunities to identify new chemical entities through structure-based drug design. To enable the determination of high-resolution structures of GPCRs, most receptors have to be engineered to overcome intrinsic hurdles such as their poor stability and low expression levels. In recent years, multiple engineering approaches have been developed to specifically address the technical difficulties of working with GPCRs, which are now beginning to make more challenging receptors accessible to detailed studies. Importantly, successfully engineered GPCRs are not only valuable in x-ray crystallography, but further enable biophysical studies with nuclear magnetic resonance spectroscopy, surface plasmon resonance, native mass spectrometry, and fluorescence anisotropy measurements, all of which are important for the detailed mechanistic understanding, which is the prerequisite for successful drug design. Here, we summarize engineering strategies based on directed evolution to reduce workload and enable biophysical experiments of particularly challenging GPCRs.https://www.mdpi.com/1420-3049/26/5/1465G protein-coupled receptorsdirected evolutionprotein engineeringNK<sub>1</sub>RNTS<sub>1</sub>RPTH<sub>1</sub>R |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yann Waltenspühl Janosch Ehrenmann Christoph Klenk Andreas Plückthun |
spellingShingle |
Yann Waltenspühl Janosch Ehrenmann Christoph Klenk Andreas Plückthun Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies Molecules G protein-coupled receptors directed evolution protein engineering NK<sub>1</sub>R NTS<sub>1</sub>R PTH<sub>1</sub>R |
author_facet |
Yann Waltenspühl Janosch Ehrenmann Christoph Klenk Andreas Plückthun |
author_sort |
Yann Waltenspühl |
title |
Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies |
title_short |
Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies |
title_full |
Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies |
title_fullStr |
Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies |
title_full_unstemmed |
Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies |
title_sort |
engineering of challenging g protein-coupled receptors for structure determination and biophysical studies |
publisher |
MDPI AG |
series |
Molecules |
issn |
1420-3049 |
publishDate |
2021-03-01 |
description |
Membrane proteins such as G protein-coupled receptors (GPCRs) exert fundamental biological functions and are involved in a multitude of physiological responses, making these receptors ideal drug targets. Drug discovery programs targeting GPCRs have been greatly facilitated by the emergence of high-resolution structures and the resulting opportunities to identify new chemical entities through structure-based drug design. To enable the determination of high-resolution structures of GPCRs, most receptors have to be engineered to overcome intrinsic hurdles such as their poor stability and low expression levels. In recent years, multiple engineering approaches have been developed to specifically address the technical difficulties of working with GPCRs, which are now beginning to make more challenging receptors accessible to detailed studies. Importantly, successfully engineered GPCRs are not only valuable in x-ray crystallography, but further enable biophysical studies with nuclear magnetic resonance spectroscopy, surface plasmon resonance, native mass spectrometry, and fluorescence anisotropy measurements, all of which are important for the detailed mechanistic understanding, which is the prerequisite for successful drug design. Here, we summarize engineering strategies based on directed evolution to reduce workload and enable biophysical experiments of particularly challenging GPCRs. |
topic |
G protein-coupled receptors directed evolution protein engineering NK<sub>1</sub>R NTS<sub>1</sub>R PTH<sub>1</sub>R |
url |
https://www.mdpi.com/1420-3049/26/5/1465 |
work_keys_str_mv |
AT yannwaltenspuhl engineeringofchallenginggproteincoupledreceptorsforstructuredeterminationandbiophysicalstudies AT janoschehrenmann engineeringofchallenginggproteincoupledreceptorsforstructuredeterminationandbiophysicalstudies AT christophklenk engineeringofchallenginggproteincoupledreceptorsforstructuredeterminationandbiophysicalstudies AT andreaspluckthun engineeringofchallenginggproteincoupledreceptorsforstructuredeterminationandbiophysicalstudies |
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