How bacterial pathogens "sweeten" host proteins to avoid immunological responses: structural study of sugar transfer by bacterial virulence factors
Lead Research Organisation:
University of East Anglia
Department Name: Graduate Office
Abstract
Antimicrobial resistance is one of the biggest threats to global health. To reduce it, we must ensure that antibiotics are used only where appropriate. Accordingly, other non-antibiotic approaches are welcome, and a better understanding of the biology of bacterial infections is required. Pathogens such as Salmonella and E. coli inject virulence factors (effectors) to suppress antimicrobial host responses, to promote colonisation. The NleB effector is highly conserved among pathogens. It transfers GlcNAc sugar residues to host proteins (GAPDH, FADD, TRADD), a "sweet tag" that inhibits NF-kB activation and apoptosis of infected cells, blocking major antimicrobial host responses.
In this project, we will use advanced NMR spectroscopy, molecular modelling (protein-ligand docking, long molecular dynamics simulations), and other biophysical techniques (ITC) to unveil the structural features at atomic detail of the molecular recognition of host proteins by the bacterial effectors NleB1, SseK1, and SseK2. Members of this conserved family modify different host proteins and exhibit distinct modes of action to suppress host responses, but, interestingly, they differ only in a very small number of amino acids. We will try to rationalise the molecular basis of their exquisite selectivity, and the impact of sugar transfer on the interactions with other host proteins such as TRAF2.
In this project, we will use advanced NMR spectroscopy, molecular modelling (protein-ligand docking, long molecular dynamics simulations), and other biophysical techniques (ITC) to unveil the structural features at atomic detail of the molecular recognition of host proteins by the bacterial effectors NleB1, SseK1, and SseK2. Members of this conserved family modify different host proteins and exhibit distinct modes of action to suppress host responses, but, interestingly, they differ only in a very small number of amino acids. We will try to rationalise the molecular basis of their exquisite selectivity, and the impact of sugar transfer on the interactions with other host proteins such as TRAF2.
People |
ORCID iD |
Thomas Hicks (Student) |
Publications
Wallace M
(2019)
Self-Correcting Method for the Measurement of Free Calcium and Magnesium Concentrations by 1H NMR.
in Analytical chemistry
Wu H
(2021)
Fucosidases from the human gut symbiont Ruminococcus gnavus.
in Cellular and molecular life sciences : CMLS
García-García A
(2021)
NleB/SseK-catalyzed arginine-glycosylation and enteropathogen virulence are finely tuned by a single variable position contiguous to the catalytic machinery.
in Chemical science
García-García A
(2021)
FUT8-Directed Core Fucosylation of N-glycans Is Regulated by the Glycan Structure and Protein Environment.
in ACS catalysis
Latorre-Muro P
(2021)
Self-acetylation at the active site of phosphoenolpyruvate carboxykinase (PCK1) controls enzyme activity.
in The Journal of biological chemistry
Martin KC
(2021)
Fucosyltransferase-specific inhibition via next generation of fucose mimetics.
in Chemical communications (Cambridge, England)
Description | I have co-authored a paper that shows we were able to change the target of SseK1, a protein related to my project, with only a single mutation. This mutation makes the protein able to target a protein called 'FADD' which the non-mutated Ssek1 cannot target. |
Exploitation Route | The work done in this project will directly contribute to understanding how they protein families, NleB and SseK, function. NleB and SseK assist E.Coli and S.Enterica respectively to suppress the innate immune system. These proteins are glycosyltransferases and are important as they are uniquely able to target the amino acid Arginine. This is unusual as typically proteins of this nature can only target the amino acids serine, threonine and asparagine. Given that these proteins are unique the work done as part of this project form the foundation of understanding how these proteins are uniquely able to target arginine. |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
Description | Collaboration with the Barbara Richichi Lab of the University of Florence |
Organisation | University of Florence |
Country | Italy |
Sector | Academic/University |
PI Contribution | Performed Nuclear Magnetic Resonance experiments. |
Collaborator Contribution | Provision of enzymes and bespoke ligands for Nuclear Magnetic Resonance and performed other biophysical experiments. |
Impact | Chemical Communications publication: http://dx.doi.org/10.1039/D0CC04847J |
Start Year | 2020 |
Description | Collaboration with the Matthew Wallace Lab of the University of East Anglia |
Organisation | University of East Anglia |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Performed Molecular Dynamics simulation. |
Collaborator Contribution | Performed NMR experiments and methodological development |
Impact | Anal. Chemistry publication http://dx.doi.org/10.1021/acs.analchem.9b03008 |
Start Year | 2018 |
Description | Collaboration with the Nathalie Juge Lab of the Quadram Institute |
Organisation | Quadram Institute Bioscience |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Performed Nuclear Magnetic Resonance experiments and Molecular Dynamics simulation. |
Collaborator Contribution | Provision of enzymes for Nuclear Magnetic Resonance and performed other biophysical experiments. |
Impact | Publication to Cellular and Molecular Life Science https://link.springer.com/article/10.1007/s00018-020-03514-x Multidisciplinary Collaboration : ITC, X-ray crystallography, Mass Spec, NMR, MD |
Start Year | 2018 |
Description | Collaboration with the Ramon Hurtado Lab of the University of Zaragoza |
Organisation | University of Zaragoza |
Country | Spain |
Sector | Academic/University |
PI Contribution | As part of this collaboration I perform Nuclear Magnetic Resonance and Molecular Dynamics simulations on the NleB and SseK enzymes. |
Collaborator Contribution | My collaborator provided enzyme samples for use with Nuclear Magnetic Resonance experiments and provided contributions to interpretation of Molecular Dynamics simulation. In addition, the Hurtado lab performed Isothermal Calorimetry Titration and X-ray diffraction experiments to compliment the structural information found from Nuclear Magnetic Resonance and Molecular Dynamics simulations. |
Impact | Publications to ACS catalysis: https://doi.org/10.1039/D1SC04065K https://doi.org/10.1021/acscatal.1c01698 Multidisciplinary Collaboration : ITC, X-ray crystallography, NMR, MD, Synthetic Chemistry |
Start Year | 2018 |
Description | Norwich Science Festival |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | The Norwich science festival is an annual event held in the city centre. I worked at the event for UEA school of pharmacy. |
Year(s) Of Engagement Activity | 2019 |
URL | https://norwichsciencefestival.co.uk/about-us/past-norwich-science-festivals/norwich-science-festiva... |