Structure determination of the 7-helix transmembrane protein receptor pSRII by solution NMR
Lead Research Organisation:
University of Cambridge
Department Name: Biochemistry
Abstract
Most living organisms are assembled from a very large number of cells. Each of these cells is a specialist that contributes through a large number of activities to the support of an organism. Cells are composed mostly of a liquid interior that accommodates the precious machinery including the building blocks of life and all the information to produce proteins. A water insoluble protective lipid layer called a membrane surrounds the cell. To function properly and also when necessary to adjust the interior workings of the cell to changing external demands the cell needs to be continuously supplemented with nutrients and instructions that come from the outside. To help with this, a large number of molecules, called membrane proteins such as e.g. ligand receptors are embedded in the lipid layer, where they span the membrane. They are the liaison centres of the cell and communicate between interior and exterior worlds to shuttle information and material across the cell-interface. Their involvement in so many complex information transfer processes and their implication in a large number of diseases have made them prime targets for the majority of existing drugs on the market. Biologists and chemists study the properties of such molecules in order to better understand the cellular and biomolecular processes. One important aspect is to understand the three-dimensional organization of these molecules. While this can be obtained quite routinely and with great success for water-soluble molecules, it is much more difficult to achieve for membrane embedded proteins using the traditional structure determination techniques of X-ray crystallography and nuclear magnetic resonance (NMR). Consequently structure determination of insoluble membrane proteins is still in its early stages. Our proposal aims at the development of the NMR technique to solve the structure of such membrane proteins. We concentrate our studies on a particular member, for us a model system that belongs to a family of proteins with many analogies to receptors. The latter have proven particularly evasive to structural studies so far and our research efforts will improve this situation.
Technical Summary
A multitude of problems make structural studies of membrane proteins a substantial challenge. Membrane proteins play key functional roles and their dysfunction is directly involved in many diseases. Elucidating the structure of membrane proteins is invaluable for understanding their function. Given the extensive difficulties, encouraging advances have been made recently using X-ray crystallography and NMR in solution and solid. Solution-state NMR spectroscopy is rapidly winning importance in the study of structure and dynamics of integral membrane proteins but so far has primarily concentrated on the more accessible b-barrel porin proteins with only few smaller a-helical proteins studied. We will study a helical transmembrane protein as a representative of a wider group of key membrane protein receptors. Our immediate goal is to determine the structure of the seven-helix transmembrane protein pSRII solubilized in a detergent micelle. We will concentrate on the development of the NMR methodology required to study such large protein-detergent systems (50-70 kDa). The feasibility of our approach is backed by our recent breakthrough results achieved over the last 12 months where we show the complete sequential assignment of this 7TM receptor. In addition, we will map the interaction of pSRII with its downstream transducer HtrII. Our techniques will be generally applicable to the study of other helical membrane proteins in solution. We will use the full potential of NMR and study protein dynamics and modes of detergent interaction and assess how these are governed by detergent choice. How far the properties of functional membrane proteins in their natural lipid environment can be correctly reproduced in detergent micelles has been the focus of a longstanding debate. Based on structure and dynamics we will be able to compare our findings with the results on pSRII available from X-ray and solid-state NMR. All our objectives will be met within the grant period of 36 months.
People |
ORCID iD |
| Daniel Nietlispach (Principal Investigator) |
Publications
Tan YL
(2018)
Characterization of Denatured States and Reversible Unfolding of Sensory Rhodopsin II.
in Journal of molecular biology
Tan Y
(2019)
Characterisation of denatured states of sensory rhodopsin II by solution-state NMR
in Journal of Molecular Biology
Oleszczuk M
(2010)
Advances in biological NMR circa WWMR 2010 in Florence.
in Journal of magnetic resonance (San Diego, Calif. : 1997)
Nietlispach D
(2011)
Solution NMR studies of polytopic a-helical membrane proteins.
in Current opinion in structural biology
Jansma AL
(2010)
NMR analysis of the structure, dynamics, and unique oligomerization properties of the chemokine CCL27.
in The Journal of biological chemistry
Ikeya T
(2010)
NMR protein structure determination in living E. coli cells using nonlinear sampling.
in Nature protocols
Hopper JT
(2013)
Detergent-free mass spectrometry of membrane protein complexes.
in Nature methods
Holland DJ
(2011)
Fast multidimensional NMR spectroscopy using compressed sensing.
in Angewandte Chemie (International ed. in English)
Gautier A
(2012)
Solution NMR studies of integral polytopic a-helical membrane proteins: the structure determination of the seven-helix transmembrane receptor sensory rhodopsin II, pSRII.
in Methods in molecular biology (Clifton, N.J.)
Gautier A
(2010)
Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy.
in Nature structural & molecular biology
De Biasio A
(2018)
A generalized approach for NMR studies of lipid-protein interactions based on sparse fluorination of acyl chains.
in Chemical communications (Cambridge, England)
Crick DJ
(2015)
Integral membrane protein structure determination using pseudocontact shifts.
in Journal of biomolecular NMR
Bostock MJ
(2019)
The role of NMR spectroscopy in mapping the conformational landscape of GPCRs.
in Current opinion in structural biology
Bostock MJ
(2012)
Compressed sensing reconstruction of undersampled 3D NOESY spectra: application to large membrane proteins.
in Journal of biomolecular NMR
Bostock M
(2012)
Recent Developments in Biomolecular NMR
| Description | Membrane proteins with seven _-helices are particularly difficult to study. Only little structural information on them is available. This is very unfortunate since many of these proteins are highly abundant. Located on the cell-surface as receptors they transfer extracellular signals across the membrane boundary into the cell interior. Involved in a multitude of signaling pathways they play a central role in many physiological processes. Their malfunction in particular is related to many forms of disease. Displayed on the cell surface, it makes them attractive targets for drug treatment thus gathering structural information would help to understand how these proteins function and how to guide future drug development. Currently, the majority of structural information relies on X-ray crystallography, which captures proteins in a static form. NMR spectroscopy on the other hand is a highly complementary technique to X-ray and it is able to uniquely provide information on dynamics as well as structure in solution. The obstacles with NMR studies of membrane proteins are the need for isotopically labeled samples and rising difficulties to achieve full structural studies of proteins with a molecular weight larger 40 kDa. Here the situation is aggravated by the need to solubilize these proteins using membrane mimics such as detergent micelles, which increases the overall size. This work investigated into the possibility to use NMR spectroscopy for a full structural study of seven-helical proteins. Using sensory rhodopsin (pSRII) as a test system, a microbial seven-helical protein receptor, our research shows that by applying our methodologies NMR can now be used to study such proteins in their functional state. Our work represents a proof of principle indicating that provided adequate samples can be prepared other 7TM proteins of comparable size, such as many class A GPCR receptors might become amenable to similar NMR studies. Encouragingly, the quality of the data indicates similar studies using even larger protein systems to be feasible. To succeed we optimized protein expression to make a range of suitably labeled NMR samples. We developed an NMR strategy, which resulted in a high-resolution NMR structure of functional pSRII; a first for a seven-helical membrane protein solved by solution-NMR spectroscopy. The quality of the structure is excellent and was validated against X-ray structures of the same protein. The protein dynamics were studied using NMR relaxation methods. These indicate that the helices are well structured but that inter-helix loop regions and residues near the bound chromophore co-factor show dynamics on different timescales. We are able to measure dynamics of backbone and side chains. We mapped protein-detergent interactions. The protein is surrounded by a torus of detergent molecules that covers the exterior of the hydrophobic sidechains. We assess the impact that differing protein environments have for NMR and protein stability. Further, we developed means to allow NMR structural studies to take place in more realistic membrane mimics, such as bicelles and nanodiscs. This will benefit studies of less robust membrane proteins. pSRII binds to a transducer as part of its signaling function and we mapped the binding interface to its partner protein. |
| Exploitation Route | Pharmaceutical research and small company bioscience businesses involved in drug development directly benefit from our proof of principle study and will be able to capitalize from our research. Primarily, our studies gather scientific data on membrane protein structure, dynamics and binding to other molecules. This knowledge is gathered to further mechanistic insight into the diverse cellular regulatory mechanisms involving membrane proteins. Using NMR we are addressing particular problems, which are difficult to pursue by X-ray crystallography (i.e. a main structural tool) but where atomic resolution information is necessary or where the results of NMR studies are highly complementary. Our research is into generating quality, world-leading research results to enhance the knowledge of biochemistry at the level of molecular processes. Our work furthers academic understanding and is relevant for the advancement of international science. More specifically in this project: Only insufficient structural information on the abundant family of seven-helical membrane proteins is available. Consequently, exploiting the use of all available structural methods that can provide information at atomic resolution is important. X-ray crystallography is the main structural technique. However, NMR spectroscopy is a complementary technique to X-ray crystallography since next to structural information it can provide information on dynamics, ligand binding, conformational changes at atomic resolution that no other high resolution technique is able to do. Through our research we have developed a strategy and methodology, which makes the study of seven-helical proteins accessible to NMR spectroscopy. The methodology employed to do such studies has been made available by us in the literature and was widely disseminated during a number of international conferences. The methodologies can be applied by other research environments, which have access to the specialized NMR equipment and are experienced with studying large size proteins. While microbial proteins such as the one studied by us are easier to handle, studies of G protein-coupled receptors (GPCRs) the largest family of eukaryotic proteins are a bigger challenge. These proteins are of significant physiological relevance as they function as relays for signal transmission into the cell. Central to many cellular processes they are also involved in a multitude of diseases and thus as drug targets become of medical and therapeutic relevance. A successful drug design benefits from structural information as a source that contributes towards unraveling how the molecular mechanisms of action work. Our study indicates that NMR can now be used to gather information on such difficult systems. Unfortunately, structural studies of GPCR proteins are still more difficult to achieve on a regular basis, as these proteins are less stable, express in small quantities and are believed also to be mobile. The extensive dynamics displayed are thought to be crucial to facilitate function, where these proteins interconvert between various states that have different activities. Influenced through binding to different ligands this results in the moderation of the downstream responses. NMR spectroscopy has the unique ability to report on the dynamical behavior of GPCRs and on ligand binding. In the near future, the study of mobility and changes in dynamics of various GPCRs upon ligand binding will contribute towards a detailed mechanistic understanding of how these molecules work. Additional molecular information on ligand binding will also be obtained from NMR spectroscopy. It is important to understand where the ligands are binding and how the protein structures are changed. This will help to direct the development of potential drugs that aim at influencing the activity of such proteins. While GPCRs tend to bind to a range of ligands, which result in different downstream effects, intention is to develop drugs that lead to a selective rather than broadband response. Influence of potential drug candidates on structure and dynamics and how the equilibria between different protein conformations are affected will be obtained from NMR studies, thus providing priceless complementary information to push ahead the drug development with the aim to combat disease. Out of this a new generation of drugs will be developed which will make better potential use of the easy accessibility of these proteins on the cell surface. While academic research, pharmaceutical and small bioscience companies are the ones immediately benefitting from the potential of our proof of principle study, the long term effects via development of drugs and a better understanding of cell signaling at a molecular level will reach much further: they will be the fight of disease and an improvement of health of the general public. Further research has been conducted into using pSRII as a model membrane protein to study protein folding. Using detergents as membrane mimetic we have used a range of biophysical techniques and NMR spectroscopy to optain a high resolution picture of the folding behaviour of pSRII. Our findings show that pSRII is folding/unfolding according to a new paradigm that is situated between the two-state folding model followed by bacteriorhodopsin and the core forming folding model followed by rhodopsin. pSRII contains features of both folding routes. Accordingly pSRII is a suitable model system to pursue further folding studies of polytopic a-helical membrane proteins. |
| Sectors | Pharmaceuticals and Medical Biotechnology |
| URL | https://doi.org/10.1016/j.jmb.2019.04.039 |
| Description | Our research has enabled the use of NMR spectroscopy in a field of structural biology, which is very underdeveloped and was thought not to be feasible prior to our efforts. Our work was initially delivered in form of a proof of principle study and was highly acclaimed in its reception, with responses and interest from academic research and pharmaceutical companies remaining strong throughout the grant period. Our methods are the foundation for further NMR studies of seven-helical membrane proteins involved in regulation of cellular processes and disease e.g. G protein-coupled receptors. Publication of our major results was accompanied by a 'Research Highlights' in Nature Methods and has resulted in the dissemination of our results through the PI as an invited speaker at over ten major international conferences in the fields of membrane protein biology, structural biology and NMR spectroscopy. Three invited, peer-reviewed reviews have further resulted from this (one published, one in press, one accepted for publication). Next to advancement in the areas of membrane protein expression and NMR methodological developments this has also lead to the expansion into novel NMR data recording and processing methodologies. The latter are universally applicable and are leading to generally substantially increased performance of the NMR equipment. We advanced academic research. We created training opportunities on national/international level. We triggered technical development in new areas. We attracted the interest of pharmaceutical research, which in longer term will lead to R&D investment. While our results are not immediately commercially exploitable they are relevant for drug research and via novel drug based therapies will improve public health. This will lead to economic prosperity and higher economical competitiveness. |
| First Year Of Impact | 2009 |
| Sector | Cultural |
| Description | Expression of G protein-coupled receptors in E.coli. |
| Amount | £63,000 (GBP) |
| Funding ID | RG53842 |
| Organisation | University of Cambridge |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 10/2010 |
| End | 10/2013 |
| Description | Expression of the b1-adrenergic receptor using Pichia pastoris. |
| Amount | £83,000 (GBP) |
| Funding ID | RG53842 |
| Organisation | University of Cambridge |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 10/2011 |
| End | 10/2014 |
| Description | Replacement of NMR Spectrometer Consoles - an Upgrade to Secure the Future Operation of the Biomolecular NMR Facility in the Department of Biochemistry, University of Cambridge |
| Amount | £22,070 (GBP) |
| Organisation | University of Cambridge |
| Department | Isaac Newton Trust |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 04/2010 |
| End | 04/2011 |
| Description | Replacement of NMR Spectrometer Consoles - an Upgrade to Secure the Future Operation of the Biomolecular NMR Facility in the Department of Biochemistry, University of Cambridge |
| Amount | £22,070 (GBP) |
| Organisation | University of Cambridge |
| Department | Isaac Newton Trust |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 05/2010 |
| End | 05/2011 |
| Description | Royal Society International Exchange Scheme |
| Amount | £12,000 (GBP) |
| Funding ID | IE161563 |
| Organisation | The Royal Society |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 03/2017 |
| End | 03/2020 |
| Description | Studies of helical membrane proteins using NMR spectroscopy |
| Amount | £62,000 (GBP) |
| Funding ID | RG53842 |
| Organisation | University of Cambridge |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 04/2009 |
| End | 04/2012 |
| Description | Upgrade of NMR Spectrometer Consoles to Secure the Future Operation of the Biomolecular NMR Facility in the Department of Biochemistry, University of Cambridge |
| Amount | £500,000 (GBP) |
| Funding ID | 094234/Z/10/Z |
| Organisation | Wellcome Trust |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 05/2011 |
| End | 05/2014 |
| Title | Compressed Sensing data reconstruction methodology |
| Description | Development of data processing methodology to accelerate NMR spectroscopy. |
| Type Of Material | Improvements to research infrastructure |
| Year Produced | 2013 |
| Provided To Others? | Yes |
| Impact | The developed methodology enables the study of more complex biological studies using NMR spectroscopy. |
| Title | NMR data processing software for the reconstruction of undersampled data |
| Description | 3D and 4D NMR data takes a long time to record. We show that a combination of sparse data sampling combined with the use of Compressed Sensing processing can lead to large time savings or lead to higher sensitivity and resolution per unit time. This boosts the performance of NMR equipment and becomes an indispensable tool for the study of large molecular weight biomolecules. The processing routines are now available as a standalone platform independent package. |
| Type Of Material | Computer model/algorithm |
| Year Produced | 2017 |
| Provided To Others? | Yes |
| Impact | Software for NMR data processing to allow reconstruction of non-uniformly sampled data. The algorithm will enter the annual contest for data processing of NUS NMR data and will be one of the leading softwares. |
| Description | Investigating the light activation mechanism of the signal receptor pSRII |
| Organisation | Academia Sinica |
| Department | Institute of Astronomy and Astrophysics |
| Country | Taiwan, Province of China |
| Sector | Academic/University |
| PI Contribution | We are making 2H,15N and 15N isotopically labeled protein samples for NMR studies of the protein in detergent micelles. In addition we are preparing samples with the receptor reconstituted into lipid nanoparticles. |
| Collaborator Contribution | Our collaborators have conducted initial investigations into the kinetics of the light cycle and on steady state light excitation. These investigations are key for our ongoing light activation study by NMR, where we are exciting the protein in-situ via laser light activation. Our collaborators have optimized techniques to improve the light excitation. THis is a key step of our investigations as the concentrated protein samples absorb a lot of the light and it has proven difficult to excite sufficient receptor without bleaching the ground state. |
| Impact | This study will see some dramatic progress during 2018. Subsequently we will publish the outcomes of our measurements. |
| Start Year | 2016 |
| Description | Investigating the light activation mechanism of the signal receptor pSRII |
| Organisation | National Tsing Hua University (Taiwan) |
| Country | Taiwan, Province of China |
| Sector | Academic/University |
| PI Contribution | We are making 2H,15N and 15N isotopically labeled protein samples for NMR studies of the protein in detergent micelles. In addition we are preparing samples with the receptor reconstituted into lipid nanoparticles. |
| Collaborator Contribution | Our collaborators have conducted initial investigations into the kinetics of the light cycle and on steady state light excitation. These investigations are key for our ongoing light activation study by NMR, where we are exciting the protein in-situ via laser light activation. Our collaborators have optimized techniques to improve the light excitation. THis is a key step of our investigations as the concentrated protein samples absorb a lot of the light and it has proven difficult to excite sufficient receptor without bleaching the ground state. |
| Impact | This study will see some dramatic progress during 2018. Subsequently we will publish the outcomes of our measurements. |
| Start Year | 2016 |
| Description | NMR analysis of the structure, dynamics, and unique oligomerization properties of the chemokine CCL27. |
| Organisation | University of California, San Diego (UCSD) |
| Department | Skaggs School of Pharmacy and Pharmaceutical Sciences |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Determination of the structure of a small novel chemokine ccl27 and study of the oligomerization behaviour of this protein which has functional relevance. Chemokines interact with 7-helical GPCRs. |
| Collaborator Contribution | Collaborator provided expression constructs and protocols for protein expression and purification |
| Impact | 1 Publication. doi:10.1074/jbc.M109.091108 This collaboration is multi-disciplinary. Biology Analytical Spectroscopy |
| Start Year | 2009 |
| Title | NMR data processing: Compressed sensing to improve NMR performance |
| Description | 3D and 4D NMR data takes a long time to record. We show that a combination of sparse data sampling combined with the use of Compressed Sensing processing can lead to large time savings or lead to higher sensitivity and resolution per unit time. This boosts the performance of NMR equipment and becomes an indispensable tool for the study of large molecular weight biomolecules. The processing algorithms are now available as a GUI supported platform independent software package. The code allows FT, CS, ME reconstruction of fully or NUS sampled data, including RQD. |
| Type Of Technology | New/Improved Technique/Technology |
| Year Produced | 2017 |
| Impact | Software for NMR data reconstruction. |
| Description | Academia Sinica talk 2012 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Institute talk, invited Academia Sinica, Taipei, Taiwan Solution NMR studies of a-helical membrane proteins Invited Speaker 3 December 2012 Was approached to contribute in workshops planned for further years |
| Year(s) Of Engagement Activity | 2012 |
| Description | Biophysics meeting 2014 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Type Of Presentation | keynote/invited speaker |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | British biophysical society conference 2014 New frontiers in biophysics University of Warwick Invited speaker title of talk: Funtional studies from 7TN receptors: insight from solution NMR spectroscopy 9-11th July 2014 Triggered stimulating discussions at the end of the session. Approached for collaboration with UK academics |
| Year(s) Of Engagement Activity | 2014 |
| Description | Cambridge NMR Conference 2014 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Type Of Presentation | workshop facilitator |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Co-organiser of the meeting: including organisation and co-responsibility for putting scientific program together NMR in structural biology Co-organiser 10-11 April 2014 University of Cambridge, Department of Chemistry Received very positive echo from audience on the quality of the scientific program that had been put together. Showed that short meetings can be excellent medium for scientific exchange. |
| Year(s) Of Engagement Activity | 2014 |
| URL | http://www-keeler.ch.cam.ac.uk/NMRDG/ |
| Description | ENC meeting 2012 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other academic audiences (collaborators, peers etc.) |
| Results and Impact | Invited speaker Compressed sensing reconstruction of undersampled NOESY spectra: application to large membrane proteins. ENC 2012: 53rd Experimental Nuclear Magnetic Resonance Conference, Miami After the talk, people approached with the intention to collaborate and also contribute to future review article. |
| Year(s) Of Engagement Activity | 2012 |
| Description | ETH talk 2014 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other academic audiences (collaborators, peers etc.) |
| Results and Impact | Invited Seminar speaker ETH Zurich, Switzerland title of talk: Structure and function of 7-helical membrane proteins studied by NMR spectroscopy 18 March 2014 talk triggered long discussion and many questions Undergraduate students found talk a very useful introduction into the subject |
| Year(s) Of Engagement Activity | 2014 |
| Description | Gottingen talk 2014 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Invited speaker. Title: Functional studies of 7tm receptors: insight from solution NMR spectroscopy SFB 803 Symposium, Georg-August University, Gottingen 29.9-1.10 2014 Gottingen Germany Was approached for collaborations following talk |
| Year(s) Of Engagement Activity | 2014 |
| Description | Leipzig talk 2013 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Postgraduate students |
| Results and Impact | Institute seminar talk University of Leipzig title of talk: Insight from NMR spectroscopy on functional studies of 7TM receptors: a progress report 29 September 2013 Talk resulted in further discussion that led to ongoing collaboration (University of Leipzig, Prof Dr D. Huster) |
| Year(s) Of Engagement Activity | 2013 |
| Description | Luebeck meeting 2014 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Type Of Presentation | keynote/invited speaker |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Invited speaker: Annual meeting of the German Biophysical society (DGFB) 14-17 September, Lubeck, Germany title of talk: Functional studies of 7TM receptors: insight from solution NMR spectroscopy Raise interest in topics that can be addressed by NMR |
| Year(s) Of Engagement Activity | 2014 |
| Description | NMR studies of a 7-helix transmembrane receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Seminar series: Biophysics talks, Chemistry Department, University of Cambridgei Invited talk, oral presentation. We show NMR spectroscopy to be well suited to solve structure of seven-helical membrane proteins. no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | NMR studies of a 7-helix transmemebrane receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Undergraduate students |
| Results and Impact | 10th CCPN UK meeting in Ambleside Invited speaker, oral presentation. Overview over how NMR can be used to study structure and function of helical membrane proteins. no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | Novartis talk 2012 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Invited talk Novartis NIBR, Drug Discovery group title: Structure determination of helical membrane proteins by NMR spectroscopy in solution 31 January 2012 This talk was the start of a very productive collaboration |
| Year(s) Of Engagement Activity | 2012 |
| Description | Solution-NMR Structure Determination of the 7-TM Receptor Sensory Rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Gordon Research Conference on Ligand Recognition and Molecular Gating, Il Ciocco Italy Invited speaker, oral presentation. We show the first determination of the structure of a microbial seven-helix membrane protein by NMR. no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | Solution-NMR structural studies of the 7-helix transmembrane receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Keystone Symposia, Santa Fe. Frontiers of NMR in Biology Invited speaker, oral presentation (45min) no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2009 |
| Description | Solution-NMR structural studies of the 7-helix transmembrane receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Type Of Presentation | keynote/invited speaker |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Biophysics seminar series at the Biozentrum - Invited speaker, oral presentation. NMR spectroscopy is complementary to X-ray crystallography and has now been shown to be able to solve structures of seven-helical membrane proteins. A detailed investigation on sensory rhodopsin is shown. no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2009 |
| Description | Solution-NMR structure determination of the 7TM receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Invited speaker, oral presentation (45min) International Magnetic Resonance Conference no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | Solution-NMR structure determination of the 7TM receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Invited speaker, oral presentation: NMR can be used now to study membrane proteins and allows to obtain structures of 7-helical bacterial proteins Invited speaker, oral presentation (45min) International Conference on Magnetic Resonance in Biological Systems no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | Structure determination of helical membrane proteins by NMR spectroscopy in solution |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Bio-NMR Foresight meeting in Bad Homburg Invited speaker, Oral presentation: How to use NMR to solve structures of polytopic membrane proteins no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2011 |
| Description | Structure determination of helical membrane proteins by NMR spectroscopy in solution |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Industry/Business |
| Results and Impact | One hour invited lecture, followed by research hearing at Novartis. Oral presentation: Overview of the role of NMR spectroscopy in the structural study of membrane proteins no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2012 |
| Description | Structure determination of the helical 7-transmembrane receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Annual meeting of the NMR society of Japan Invited speaker, oral presentation: The structural study of a 7 helical membrane protein using NMR spectroscopy. NMR of larger proteins. no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | The solution-NMR structure determination of the 7TM receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Type Of Presentation | keynote/invited speaker |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | CIC bioGUNE Center for Cooperative Research in Biosciences, Bilbao. Invited speaker, oral presentation. Structural studies of membrane proteins by X-ray crystallography are now complemented by NMR spectroscopy in solution. The two methods are highly complementary no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |
| Description | The solution-NMR structure determination of the 7TM receptor sensory rhodopsin II |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | International conference, Cold Spring Harbor Asia organisation. Location: Suzhou, China Invited speaker, oral presentation. For the first time we show how the structure of a 7-helical membrane protein was determined by NMR no actual impacts realised to date |
| Year(s) Of Engagement Activity | 2010 |