Unravelling the Bam complex
Lead Research Organisation:
University of Birmingham
Department Name: Sch of Biosciences
Abstract
The growing resistance of micro-organisms to antimicrobial therapies, such as antibiotics, is a significant global public health issue. In Europe alone this is currently estimated to result in an additional 25,000 deaths each year, approximately 2.5 million avoidable days in hospital and an economic burden estimated to be at least £1.2 Billion per year. Resistance is most serious for Gram-negative bacteria, with essentially few antibiotics under development or likely to be available for clinical use in the near future. A recent report by Professor Dame Sally Davies, the Government's Chief Medical Officer, likened it to a 'ticking time bomb' and warned that routine operations could become deadly in just 20 years if we lose the ability to fight infection.
The understanding of the Gram-negative bacterial cell envelope is critical to developing new antimicrobial agents. All Gram-negative bacteria possess two membranes that enclose the cell, separated by a space known as the periplasm. The outer of the two membranes is particularly resistant to the penetration of small molecules. The main function of this membrane is to form a semi-permeable layer that protects the bacterium from the environment but also to control the movement of molecules into and out of the cell and how the cell interacts with its environment. It is the proteins within this membrane that control these mechanisms by providing essential physiological, pathogenic and drug resistance functions and hence are the instruments of microbial warfare as they mediate many of the lethal processes responsible for infection and disease progression. Identifying compounds that can prevent the formation of this membrane could lead to the development of the next generation of antimicrobials.
Recently a single protein complex called the Bam complex was identified as being responsible for the folding and insertion of most if not all of the proteins into the outer membrane and hence has been identified as a key bottleneck in the formation of this membrane. Understanding how this complex works is therefore critical as the design of compounds that inhibit this process would inhibit outer membrane protein biogenesis and therefore essential physiological, pathogenic and drug resistance functions and could prove useful in combating diverse Gram-negative pathogens.
In this research project we plan to characterise the mechanistic processes the Bam complex undertakes in order to fold proteins into the outer membrane, we will identify ligand interaction sites and binding pockets that form during outer membrane protein insertion and thus provide valuable mechanistic insights that will aid in the discovery of molecular inhibitors and new classes of antimicrobial agents.
The understanding of the Gram-negative bacterial cell envelope is critical to developing new antimicrobial agents. All Gram-negative bacteria possess two membranes that enclose the cell, separated by a space known as the periplasm. The outer of the two membranes is particularly resistant to the penetration of small molecules. The main function of this membrane is to form a semi-permeable layer that protects the bacterium from the environment but also to control the movement of molecules into and out of the cell and how the cell interacts with its environment. It is the proteins within this membrane that control these mechanisms by providing essential physiological, pathogenic and drug resistance functions and hence are the instruments of microbial warfare as they mediate many of the lethal processes responsible for infection and disease progression. Identifying compounds that can prevent the formation of this membrane could lead to the development of the next generation of antimicrobials.
Recently a single protein complex called the Bam complex was identified as being responsible for the folding and insertion of most if not all of the proteins into the outer membrane and hence has been identified as a key bottleneck in the formation of this membrane. Understanding how this complex works is therefore critical as the design of compounds that inhibit this process would inhibit outer membrane protein biogenesis and therefore essential physiological, pathogenic and drug resistance functions and could prove useful in combating diverse Gram-negative pathogens.
In this research project we plan to characterise the mechanistic processes the Bam complex undertakes in order to fold proteins into the outer membrane, we will identify ligand interaction sites and binding pockets that form during outer membrane protein insertion and thus provide valuable mechanistic insights that will aid in the discovery of molecular inhibitors and new classes of antimicrobial agents.
Technical Summary
To elucidate the mechanistic details of Bam complex mediated protein folding, we have developed two systems:
1. A surface tethered membrane bilayer system that allows real time monitoring of structural changes within the complex during Bam mediated protein folding.
2. A styrene maleic acid lipid particle (SMALP) system that encapsulates the complex and its lipid environment and provides a framework for probing lipid composition and also provides a novel way of structure determination by electron microscopy (EM).
Using these systems we have developed the following work packages:
A. Map the folding mechanism within the membrane. Using our surface tethered bilayer system in combination with selective deuteration and neutron reflectometry we will probe real time structural changes within the complex during chaperone binding and OMP folding, whilst stalled intermediates will allow us to characterise intermediates in the folding pathway. Quartz crystal microbalance will provide details on binding affinity, kinetics and loading. Analysing mutants and component removal will allow us to identify the role each plays in the folding process.
B. Elucidate atomic snapshots of the folding pathway. Using the latest generation cryo-EM in combination with SMALPs snapshots of the Bam folding pathway will be taken. SMALPs allow simple EM imaging of membrane proteins in all dimensions and will allow us to probe the folding event by A) using rapid freezing at set points during the folding cycle and B) utilising stalled intermediates. Structures will be resolved to identify the mode by which Bam mediated membrane insertion occurs.
C. Identify whether Bam modulates the local lipid environment. SMALPs will be used to extract the Bam complex and its associated lipid and LPS. This will be analysed by mass spectrometry and compared to the bulk outer membrane. The effect of component removal will be analysed in order to address the role each plays in lipid/LPS recuitment.
1. A surface tethered membrane bilayer system that allows real time monitoring of structural changes within the complex during Bam mediated protein folding.
2. A styrene maleic acid lipid particle (SMALP) system that encapsulates the complex and its lipid environment and provides a framework for probing lipid composition and also provides a novel way of structure determination by electron microscopy (EM).
Using these systems we have developed the following work packages:
A. Map the folding mechanism within the membrane. Using our surface tethered bilayer system in combination with selective deuteration and neutron reflectometry we will probe real time structural changes within the complex during chaperone binding and OMP folding, whilst stalled intermediates will allow us to characterise intermediates in the folding pathway. Quartz crystal microbalance will provide details on binding affinity, kinetics and loading. Analysing mutants and component removal will allow us to identify the role each plays in the folding process.
B. Elucidate atomic snapshots of the folding pathway. Using the latest generation cryo-EM in combination with SMALPs snapshots of the Bam folding pathway will be taken. SMALPs allow simple EM imaging of membrane proteins in all dimensions and will allow us to probe the folding event by A) using rapid freezing at set points during the folding cycle and B) utilising stalled intermediates. Structures will be resolved to identify the mode by which Bam mediated membrane insertion occurs.
C. Identify whether Bam modulates the local lipid environment. SMALPs will be used to extract the Bam complex and its associated lipid and LPS. This will be analysed by mass spectrometry and compared to the bulk outer membrane. The effect of component removal will be analysed in order to address the role each plays in lipid/LPS recuitment.
Planned Impact
Academic Impact:
The principal beneficiaries of this research are likely to be academic researchers in numerous fields. By identifying the mechanism by which the Bam complex functions, we will enhance the UK knowledge economy and contribute to the global understanding of outer membrane biogenesis and hence microbial pathogenesis, virulence and multidrug resistance and hence will be of interest to immunologists and bacteriologists. Furthermore the Bam complex is a new target for the development of novel antimicrobials, a goal desperately needed following the emergence of bacteria that are resistant to available antibiotics, especially with essentially few antibiotics under development or likely to be available for clinical use in the near future.
More generally, uncovering the key mechanism of outer membrane protein biogenesis will be of interest to anyone interested in biological membranes, protein folding, signalling, trafficking, secretion, drug discovery and bacterial physiology.
Commercial Impact:
While we do not anticipate our research will produce commercially exploitable results immediately, in the medium term, the Bam complex represents a novel target for the development of antimicrobials. An important beneficiary would be the pharmaceutical industry, which would be given the ability to rationally design inhibitors of Bam complex function, thus providing new opportunities to attenuate bacteria in the pursuit of anti-infective agents. The emergence of bacteria that are resistant to available antibiotics represents an enormous and growing global threat, requiring new targets and strategies to combat infection. The Bam complex offers an accessible target for intervention against a variety of pathogenic Gram negative bacteria. For example, Gram-negative bacilli cause respiratory problems (Hemophilus influenzae, Pseudomonas aeruginosa), urinary problems (Escherichia coli, Proteus mirabilis), and gastrointestinal problems (Helicobacter pylori, Salmonella enteritidis) whilst Gram-negative cocci cause sexually transmitted disease such as Neisseria gonorrhoeae, and others meningitis, e.g. Neisseria meningitidis.
Societal Impact:
This proposal will impact upon society by improving our knowledge and understanding of the Gram-negative outer membrane, the essential organelle which protects all Gram-negative bacteria, with inserted proteins influencing pathogenicity, virulence and resistance. A recent report by Professor Dame Sally Davies, the Government's Chief Medical Examiner, likened antimicrobial resistance to a 'ticking time bomb' and warned that routine operations could become deadly in just 20 years if we lose the ability to fight infection. A new generation of antimicrobials against novel targets is therefore desperately required. Currently hospital acquired infections cost the NHS £1 billion a year and approximately 70% of all intensive care unit infections are the result of Gram-negative bacteria. The development of novel antibiotics is therefore likely to impact on every member of society, from those suffering from an infection, to the families of patients, carers and health professionals. We aim to uncover the key events in Bam complex function, the protein complex responsible for folding and inserting all outer membrane proteins. If these results can be exploited, the health benefits to society will be immense.
The principal beneficiaries of this research are likely to be academic researchers in numerous fields. By identifying the mechanism by which the Bam complex functions, we will enhance the UK knowledge economy and contribute to the global understanding of outer membrane biogenesis and hence microbial pathogenesis, virulence and multidrug resistance and hence will be of interest to immunologists and bacteriologists. Furthermore the Bam complex is a new target for the development of novel antimicrobials, a goal desperately needed following the emergence of bacteria that are resistant to available antibiotics, especially with essentially few antibiotics under development or likely to be available for clinical use in the near future.
More generally, uncovering the key mechanism of outer membrane protein biogenesis will be of interest to anyone interested in biological membranes, protein folding, signalling, trafficking, secretion, drug discovery and bacterial physiology.
Commercial Impact:
While we do not anticipate our research will produce commercially exploitable results immediately, in the medium term, the Bam complex represents a novel target for the development of antimicrobials. An important beneficiary would be the pharmaceutical industry, which would be given the ability to rationally design inhibitors of Bam complex function, thus providing new opportunities to attenuate bacteria in the pursuit of anti-infective agents. The emergence of bacteria that are resistant to available antibiotics represents an enormous and growing global threat, requiring new targets and strategies to combat infection. The Bam complex offers an accessible target for intervention against a variety of pathogenic Gram negative bacteria. For example, Gram-negative bacilli cause respiratory problems (Hemophilus influenzae, Pseudomonas aeruginosa), urinary problems (Escherichia coli, Proteus mirabilis), and gastrointestinal problems (Helicobacter pylori, Salmonella enteritidis) whilst Gram-negative cocci cause sexually transmitted disease such as Neisseria gonorrhoeae, and others meningitis, e.g. Neisseria meningitidis.
Societal Impact:
This proposal will impact upon society by improving our knowledge and understanding of the Gram-negative outer membrane, the essential organelle which protects all Gram-negative bacteria, with inserted proteins influencing pathogenicity, virulence and resistance. A recent report by Professor Dame Sally Davies, the Government's Chief Medical Examiner, likened antimicrobial resistance to a 'ticking time bomb' and warned that routine operations could become deadly in just 20 years if we lose the ability to fight infection. A new generation of antimicrobials against novel targets is therefore desperately required. Currently hospital acquired infections cost the NHS £1 billion a year and approximately 70% of all intensive care unit infections are the result of Gram-negative bacteria. The development of novel antibiotics is therefore likely to impact on every member of society, from those suffering from an infection, to the families of patients, carers and health professionals. We aim to uncover the key events in Bam complex function, the protein complex responsible for folding and inserting all outer membrane proteins. If these results can be exploited, the health benefits to society will be immense.
Organisations
- University of Birmingham (Lead Research Organisation)
- Rutherford Appleton Laboratory (Collaboration)
- UNIVERSITY OF LEICESTER (Collaboration)
- Science and Technologies Facilities Council (STFC) (Collaboration)
- Sanofi (Collaboration)
- UNIVERSITY OF BIRMINGHAM (Collaboration)
- University of Warwick (Collaboration)
- University of Queensland (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
- KING'S COLLEGE LONDON (Collaboration)
People |
ORCID iD |
Timothy Knowles (Principal Investigator) |
Publications
Boelter G
(2022)
The lipoprotein DolP affects cell separation in Escherichia coli, but not as an upstream regulator of NlpD
in Microbiology
Clifton LA
(2019)
Structural Investigations of Protein-Lipid Complexes Using Neutron Scattering.
in Methods in molecular biology (Clifton, N.J.)
Cooper BF
(2024)
An octameric PqiC toroid stabilises the outer-membrane interaction of the PqiABC transport system.
in EMBO reports
Cranford-Smith T
(2020)
Iron is a ligand of SecA-like metal-binding domains in vivo.
in The Journal of biological chemistry
Cranford-Smith T
(2019)
Iron is a ligand of SecA-like metal-binding domains in vivo
Hall S
(2021)
Surface-tethered planar membranes containing the ß-barrel assembly machinery: a platform for investigating bacterial outer membrane protein folding
in Biophysical Journal
Hall SCL
(2020)
Adsorption of a styrene maleic acid (SMA) copolymer-stabilized phospholipid nanodisc on a solid-supported planar lipid bilayer.
in Journal of colloid and interface science
Hughes GW
(2019)
Evidence for phospholipid export from the bacterial inner membrane by the Mla ABC transport system.
in Nature microbiology
Isom GL
(2017)
MCE domain proteins: conserved inner membrane lipid-binding proteins required for outer membrane homeostasis.
in Scientific reports
Jain N
(2018)
Minichaperone (GroEL191-345) mediated folding of MalZ proceeds by binding and release of native and functional intermediates.
in Biochimica et biophysica acta. Proteins and proteomics
Jamshad M
(2019)
The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity.
in eLife
Karunakaran MM
(2020)
Butyrophilin-2A1 Directly Binds Germline-Encoded Regions of the V?9Vd2 TCR and Is Essential for Phosphoantigen Sensing.
in Immunity
Lord S
(2024)
Uncovering the mechanisms of MuRF1-induced ubiquitylation and revealing similarities with MuRF2 and MuRF3
in Biochemistry and Biophysics Reports
Mamou G
(2022)
Peptidoglycan maturation controls outer membrane protein assembly.
in Nature
Morris FC
(2018)
YraP Contributes to Cell Envelope Integrity and Virulence of Salmonella enterica Serovar Typhimurium.
in Infection and immunity
Odintsova E
(2020)
Binding of the periplakin linker requires vimentin acidic residues D176 and E187.
in Communications biology
Pokorny L
(2024)
The vaccinia chondroitin sulfate binding protein drives host membrane curvature to facilitate fusion.
in EMBO reports
Pollock NL
(2019)
SMA-PAGE: A new method to examine complexes of membrane proteins using SMALP nano-encapsulation and native gel electrophoresis.
in Biochimica et biophysica acta. Biomembranes
Ratkeviciute G
(2021)
Methods for the solubilisation of membrane proteins: the micelle-aneous world of membrane protein solubilisation
in Biochemical Society Transactions
Salim M
(2017)
BTN3A1 Discriminates ?d T Cell Phosphoantigens from Nonantigenic Small Molecules via a Conformational Sensor in Its B30.2 Domain.
in ACS chemical biology
Description | Gram-negative bacteria, for example E. coli (gut bacteria that in some cases can cause severe disease), Neisseria gonorrhoea (Causative agent of the sexually transmitted disease gonorrhoea) and Neisseria meningitidis (causative agent of bacterial meningitis) are surrounded by two lipid membranes that make them particularly resistant to antibiotic treatment. The outer membrane makes direct contact with the environment and hence us during infection. Understanding how this membrane forms is of critical importance as it could lead to the development of novel antibiotics that are desperately needed to treat the rising trend of antimicrobial resistance. Since 2000 many of the mechanisms have been identified but how lipids get to the outer membrane has remained an enigma. The research conducted during this grant has led to the discovery of the first mechanism of phospholipid transport towards the outer membrane in gram-negative bacteria. This pathway, known as the Mla pathway, was originally believed to transport lipids away from the outer membrane but through biochemical assays developed as part of this grant we have realised the pathway can also work in the opposite direction. This research is therefore one of the final keys in understanding the biogenesis of the outer membrane in gram-negative bacteria and could lead to the discovery of new antimicrobials against gram-negative bacteria that are desperately needed to combat the growing threat of antimicrobial resistance. As part of the original proposal focusing on the Bam complex, one of the main objectives was to produce a stalled intermediate on the Bam pathway. Although our original plan to produce a stalled intermediate failed, we have recently developed a new way to stall the complex which has proven highly successful. We have also taken this research further and have found BepA, a protease responsible for removing stalled components on the folding pathway, interacts with Bam and its stalled intermediate. We are now in a position to take this further for structure elucidation. This we hope will lead to new grant capture and publications. As part of the original proposal we have also developed a new assay system for screening Bam complex assisted protein folding. This system, initially using a tethered membrane system, and now more recently a novel floating based membrane system, provides a new surface based sensor system for studying protein folding amongst other things. This work has recently been published and provides the means to further study the folding of the myriad outer membrane proteins in Gram-negative bacteria. |
Exploitation Route | Academic Our research has opened up a completely new avenue in to research of the outer membrane of bacteria. The proteins involved have homologues in all gram-negative bacteria but also eukaryotes as well, thus the results of this research has far reaching relevance. The novel methods we have developed are applicable to almost all membrane protein systems. To highlight this research we plan to present this work at international conferences, seminars and within the press. Knowles also has numerous national and international collaborations, all will benefit from the use of this technology. Pharma Our research area focuses on novel mechanisms for the biogenesis of the outer membrane in gram-negative bacteria. Thus the proteins and pathways we study could be targeted for the development of novel antimicrobials. Knowles already has or has had numerous pharma collaborators. Knowles will engage with Pharma the results of this research to further develop interactions and collaborations. Society In the long term our research could lead to the development of novel antimicrobials that could be used to face the growing threat of antimicrobial resistance. Knowles will present the findings of this research at public engagement events, e.g. Think tank, cafe scientifique, open days etc. |
Sectors | Agriculture Food and Drink Chemicals Healthcare Pharmaceuticals and Medical Biotechnology |
URL | https://www.biorxiv.org/content/10.1101/388546v1 |
Description | Diamond User Committee |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Participation in a guidance/advisory committee |
URL | http://www.diamond.ac.uk/Users/DUC.html |
Description | ISIS neutron source user committee |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Participation in a guidance/advisory committee |
URL | https://www.isis.stfc.ac.uk/Pages/User-Committee.aspx |
Description | An accurate eukaryotic plasma membrane assay for coronavirus binding |
Amount | £123,406 (GBP) |
Funding ID | BB/V01983X/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2021 |
End | 09/2021 |
Description | An in vitro gram-negative envelope mimetic: a new way to study membrane biology - BBSRC Pioneer Award |
Amount | £193,302 (GBP) |
Funding ID | BB/Y513179/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 11/2023 |
End | 10/2024 |
Description | Crossing the periplasmic void, elucidating the mechanisms of phospholipid transport in Gram-negative bacteria |
Amount | £549,517 (GBP) |
Funding ID | BB/S017283/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 08/2019 |
End | 08/2023 |
Title | Ace2 - spike surface sensor |
Description | The technology is based on sensor science. We have developed a highly accurate biological membrane mimetic on top of a sensor surface that allows the study of host pathogen interactions amongst other things. In this grant we have used the ACE2/Spike interaction attributed as the main/first interaction in COVID infection. Using this system we can, in real time, detect binding of Spike protein but also gain key structural information regarding the membrane surface and how the interactions between the two proteins occurs. This technology could be applied to any virus/host interaction, to bacterial/host interactions and any other system that utilises the membrane. Why is this system needed? Using Covid as an example, understanding the interactions between the coronavirus spike (S) protein and the mammalian ACE2 protein is crucial in the fight against SARS-CoV-2. These two membrane proteins form complex interactions with each other and other parts of the membrane, causing extensive membrane perturbation. It is the membrane environment in its entirety that is needed for complete and accurate understanding of the exact interactions between these two membrane proteins and viral entry. Whilst many may use truncated or mutated versions of these proteins or use detergents to maintain solubility and stability, these don't reflect the true nature of the proteins. This is why we have created a "true to nature" membrane mimic with the SARS-CoV-2 spike protein and the mammalian ACE2 protein, both in their full native structures, situated within a lipid membrane. By creating these membrane mimics we have used neutron reflectometry and QCM to study not only the interactions between the two membrane proteins but also the membrane environment. These techniques enable different membrane environments (i.e. different lipids compositions and additional membrane proteins (e.g. TMPRSS2, B0AT1) to be examined and their effects on viral binding and membrane rearrangement to be studied. This membrane rearrangement has been a target for antivirals against coronaviruses in the past. The MERS-CoV was prevented from entry in a pseudotype assay by competitive inhibition using a heptad repeat 2 peptide of the S protein (Gao et al. 2013) and by a 5 helix bundle, designed as a mimic of the final S fusion intermediate (Sun et al. 2017). |
Type Of Material | Technology assay or reagent |
Year Produced | 2022 |
Provided To Others? | No |
Impact | Not yet. This work is just being prepared for publication. |
Title | Surface based assay to screen the activity of the Bam complex. An essential protein complex within the gram-negative outer membrane responsible for protein folding |
Description | Surface based assay developing a tethered floating bilayer system to study bacterial protein folding including the potential to screen for compounds to inhibit activity |
Type Of Material | Technology assay or reagent |
Year Produced | 2021 |
Provided To Others? | Yes |
Impact | The current research has led to us further developing the system to allow the use of completely floating bilayers. This has recently been awarded a BBSRC Pioneer award (BB/Y513179/1). |
Title | Iron is a ligand of SecA-like metal-binding domains in vivo (dataset) |
Description | EPR data accompanying the publication. Description of the data in the .DSC and data in the .DTA Bruker files. |
Type Of Material | Database/Collection of data |
Year Produced | 2020 |
Provided To Others? | Yes |
URL | https://risweb.st-andrews.ac.uk:443/portal/en/datasets/iron-is-a-ligand-of-secalike-metalbinding-dom... |
Title | MlaC crystal structure |
Description | Protein structure |
Type Of Material | Database/Collection of data |
Year Produced | 2019 |
Provided To Others? | Yes |
Impact | Publication in Nature Microbiology. Conference presentations, outreach activities at university open days. |
Title | PqiC crystal structure |
Description | Octomeric crystal structure of PqiC |
Type Of Material | Database/Collection of data |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | Improved understanding of lipid transport processes in gram-negative bacteria |
Title | PqiC crystal structure - alternate conformation |
Description | Crystal structure of PqiC - alternate conformation |
Type Of Material | Database/Collection of data |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | Provided additional insight into phospholipid transport in gram-negative bacteria |
Description | Bam complex |
Organisation | University of Queensland |
Country | Australia |
Sector | Academic/University |
PI Contribution | Research - Protein structure analysis |
Collaborator Contribution | Research - TRADIS screening, mutagenesis, cell based assays. |
Impact | Numerous publications. |
Start Year | 2009 |
Description | DsbD |
Organisation | Imperial College London |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Protein expression, purification and characterisation. lipidic cubic phase crystallisation and sample preparation for electron microscopy. |
Collaborator Contribution | MPL - Provision of LCP plates, trainining in LCP screening. Expertise. Use of Sec-Mals Leicester - expertise in electron microscopy. Structure determination by EM. Training and equipment usage. |
Impact | No outcomes yet. |
Start Year | 2018 |
Description | DsbD |
Organisation | Rutherford Appleton Laboratory |
Department | Membrane Protein Laboratory |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Protein expression, purification and characterisation. lipidic cubic phase crystallisation and sample preparation for electron microscopy. |
Collaborator Contribution | MPL - Provision of LCP plates, trainining in LCP screening. Expertise. Use of Sec-Mals Leicester - expertise in electron microscopy. Structure determination by EM. Training and equipment usage. |
Impact | No outcomes yet. |
Start Year | 2018 |
Description | DsbD |
Organisation | University of Leicester |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Protein expression, purification and characterisation. lipidic cubic phase crystallisation and sample preparation for electron microscopy. |
Collaborator Contribution | MPL - Provision of LCP plates, trainining in LCP screening. Expertise. Use of Sec-Mals Leicester - expertise in electron microscopy. Structure determination by EM. Training and equipment usage. |
Impact | No outcomes yet. |
Start Year | 2018 |
Description | EM Bam complex |
Organisation | Sanofi |
Department | Genzyme Corporation |
Country | United States |
Sector | Private |
PI Contribution | Protein expression, purification, characterisation and activity assay. Provided Styrene maleic acid polymer free of charge for their research. |
Collaborator Contribution | Electron microscopy analysis of the protein complex produced. |
Impact | Dataset collected. Analysis not complete. |
Start Year | 2017 |
Description | Leney |
Organisation | University of Birmingham |
Department | School of Biosciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Providing expertise in Protein NMR and data analysis. Protein expression and purification. Isotope labelling. |
Collaborator Contribution | Mass spectrometry analysis of samples. |
Impact | Too early. |
Start Year | 2021 |
Description | Mla pathway |
Organisation | University of Birmingham |
Department | School of Biosciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise in small angle scattering (SAXS), training in small angle scattering. Access to purification facilities, UV spectrophotometry. Data collection at SAXS facilities (ESRF, Grenoble, France). Data analysis. |
Collaborator Contribution | Expertise in protein crystallisation. Access to protein crystallisation facilities and equipment. Crystallisation consumables. Data collection and analysis. |
Impact | Paper and grant being submitted 2018 |
Start Year | 2017 |
Description | MlaCD |
Organisation | King's College London |
Department | Randall Division of Cell & Molecular Biophysics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise in protein expression/purification, in vitro assays. Sample preparation for EM, data collection and analysis. |
Collaborator Contribution | Electron microscopy data processing. Molecular dynamic simulations |
Impact | Paper in preparation. |
Start Year | 2021 |
Description | MlaCD |
Organisation | University of Warwick |
Department | School of Life Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise in protein expression/purification, in vitro assays. Sample preparation for EM, data collection and analysis. |
Collaborator Contribution | Electron microscopy data processing. Molecular dynamic simulations |
Impact | Paper in preparation. |
Start Year | 2021 |
Description | Morris |
Organisation | University of Birmingham |
Department | College of Medical and Dental Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | NMR analysis and data interpretation. Binding affinity analysis. Equipment usage. |
Collaborator Contribution | In vivo assays, in vitro assays, western blotting, Mutagenesis. |
Impact | Paper in preparation. Estimation submission date April 2022. |
Start Year | 2021 |
Description | Muscle atrophy |
Organisation | University of Birmingham |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provided expertise, equipment usage and training in protein expression and purification. |
Collaborator Contribution | Expertise in functional assays. |
Impact | Grant application - under review. |
Start Year | 2018 |
Description | Rutherford appleton laboratory |
Organisation | Science and Technologies Facilities Council (STFC) |
Department | ISIS Neutron and Muon Source |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Research - producing samples for neutron study, developing new methods for surface deposition. Publication preparation |
Collaborator Contribution | Neutron science research Publication preparation |
Impact | Publication under review in Nature microbiology |
Start Year | 2013 |
Description | Sec Pathway |
Organisation | University of Birmingham |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | NMR and SAXS spectroscopy studies. Proof reading |
Collaborator Contribution | Experimental support Proof reading Paper writing |
Impact | Two publications currently under review A third in preparation. |
Start Year | 2016 |
Description | Willcox |
Organisation | University of Birmingham |
Department | Institute of Cancer and Genomic Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Nuclear Magnetic resonance protein structure determination Publication writing. |
Collaborator Contribution | Surface plasmon resonance studies. Publication writing |
Impact | 3 Publications |
Start Year | 2010 |
Description | Applicant visit day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Undergraduate students |
Results and Impact | The purpose of this meeting was to highlight the research being undertaken at the University of Birmingham. To get prospective students excited by the research undertaken at the University. |
Year(s) Of Engagement Activity | 2020 |
Description | British biophysical society meeting |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | This hasn't taken place yet but is planned for July 2020. |
Year(s) Of Engagement Activity | 2020 |
Description | Invited talk at conference - Examining Membrane Biochemistry with Neutron Reflectometry - UK |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Presentation at Conference |
Year(s) Of Engagement Activity | 2022 |
Description | Invited Talk at another University - Aston University 2023 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Postgraduate students |
Results and Impact | Invited Speaker for Aston University Seminar Series. |
Year(s) Of Engagement Activity | 2023 |
Description | Invited speaker - University of Kent |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Undergraduate students |
Results and Impact | Invited speaker at Kent University to approximately 50-100 students/academics. This has led to a number of new collaborations using technology I have developed at University of Birmingham. |
Year(s) Of Engagement Activity | 2018 |
Description | Invited speaker Gordon Research conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | An invited talk to a Gordon Research Conference to > 300 academics. This sparked questions and discussion afterwards and new collaborations have been initiated. |
Year(s) Of Engagement Activity | 2018 |
Description | Media Video |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Participated in a media video advertising the University of Birmingham's Henry Wellcome Building for Biomolecular NMR. |
Year(s) Of Engagement Activity | 2018 |
Description | NMSUM 2017 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Presented a seminar at the Neutron and Muon Users meeting 2017. Purpose was to highlight my research on the Bam complex and the novel methods we are using for studying it. The overall aim at this conference was to highlight how neutrons can be used to study membrane proteins. This will hopefully result in others adopting the methods I presented to study their own systems. |
Year(s) Of Engagement Activity | 2017 |
Description | Vereingung fur Allgemeine und Angewandte Mikrobiologie |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Conference - plenary speaker. No outcomes other than requests for further information. |
Year(s) Of Engagement Activity | 2019 |