Japan Partnering : Damage free structures of enzymes of denitrification pathway and their complexes using SF-ROX and SFX at SACLA XFEL
Lead Research Organisation:
University of Liverpool
Department Name: Institute of Integrative Biology
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
People |
ORCID iD |
| Samar Hasnain (Principal Investigator) |
Publications
Amporndanai K
(2020)
Novel Selenium-based compounds with therapeutic potential for SOD1-linked amyotrophic lateral sclerosis
in eBioMedicine
Eady R
(2022)
New horizons in structure-function studies of copper nitrite reductase
in Coordination Chemistry Reviews
Gopalasingam CC
(2019)
Dimeric structures of quinol-dependent nitric oxide reductases (qNORs) revealed by cryo-electron microscopy.
in Science advances
Gopalasingam CC
(2022)
Frontiers in metalloprotein crystallography and cryogenic electron microscopy.
in Current opinion in structural biology
Hedison TM
(2019)
Unexpected Roles of a Tether Harboring a Tyrosine Gatekeeper Residue in Modular Nitrite Reductase Catalysis.
in ACS catalysis
Hedison TM
(2019)
Solvent-slaved protein motions accompany proton coupled electron transfer reactions catalysed by copper nitrite reductase.
in Chemical communications (Cambridge, England)
Jamali M
(2020)
The active form of quinol-dependent nitric oxide reductase from Neisseria meningitidis is a dimer
in IUCrJ
Muench SP
(2019)
The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM.
in IUCrJ
Rose S
(2022)
Single crystal spectroscopy and multiple structures from one crystal (MSOX) define catalysis in copper nitrite reductases
in Proceedings of the National Academy of Sciences
| Description | Copper-containing nitrite reductases (CuNiRs), encoded by nirK gene, are found in all kingdoms of life with only 5% of CuNiR denitrifiers having two or more copies of nirK. Recently, we have identified two copies of nirK genes in several a-proteobacteria of the order Rhizobiales including Bradyrhizobium sp. ORS 375, encoding a 4-domain heme-CuNiR and the usual 2-domain CuNiR (Br2DNiR). Compared with two of the best-studied 2-domain CuNiRs represented by the blue (AxNiR) and green (AcNiR) sub-classes, Br2DNiR, a blue CuNiR, shows a 20-fold lower catalytic efficiency despite a sequence identity of ~70%. Advanced synchrotron radiation (SR) and X-ray free electron laser (XFEL) are used to obtain most accurate (atomic resolution with unrestrained SHELX refinement) and damage-free (free from radiation-induced chemistry) structures, in as-isolated, substrate-bound and product-bound states. This combination has shed unprecedented light on the protonation states of essential catalytic residues, new reaction intermediates and how catalytic efficiency is modulated. This work was published in Science Advances (Rose et al., Sci. Adv. 2021; 7 : eabd8523). We recently combined on-line single crystal spectrorocopy with MSOX serial crystallography to report a step change in our understanding of Nitrite catalysis (reported in PNAS 2022 https://doi.org/10.1073/pnas.2205664119. |
| Exploitation Route | UK-JAPAN sympsoium is scheduled to be held in June 2024. |
| Sectors | Agriculture Food and Drink Environment Manufacturing including Industrial Biotechology |
| Description | This partnering award has helped to strengthen the UK-Japan collaboration and has brought participation by the UK scientists and administrators with Japanese scientistis and science administration. The 2 days symposium in Liverpool in June 2024 which has attracted participation from STFC executive director os large facilitiess, Director-General of ILL (Grenoble), Director of JSPS London, President and CEO of Astex Pharmaceuticals, Vice Chancellor of the University of Liverpool and Pro-VC for Global engagement and partnership. |
| First Year Of Impact | 2024 |
| Sector | Agriculture, Food and Drink,Environment,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Cultural Societal Policy & public services |
| Description | Development of ebselen and its derivatives as a potential drug for ALS |
| Amount | $250,000 (USD) |
| Organisation | The ALS Association |
| Sector | Charity/Non Profit |
| Country | United States |
| Start | 03/2019 |
| End | 04/2020 |
| Description | Towards a paradigm shift in understanding of membrane-bound Nitric Oxide reductase and its complexes with the electron donor and NO-producing enzyme |
| Amount | £796,523 (GBP) |
| Funding ID | BB/X015491/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 11/2023 |
| End | 11/2026 |
| Title | We have developed and exploited advanced crystallographic approaches, including MSOX (Multiple strcutuures from one crystal) and SF-ROX (Serial Femtosecond oscillation crystallography) for structure-function studies of metalloenzymes |
| Description | We have pioneered the MSOX approach for constructing structural animations ('movies') to visualize redox- driven catalytic reactions and have helped advance the SF-ROX approach to provide FRIC structures using femtosecond (<10fs) X-ray pulses from XFEL at SACLA. |
| Type Of Material | Improvements to research infrastructure |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| Impact | S. L. Rose, S. V. Antonyuk, D. Sasaki, K. Yamashita, K. Hirata, Go Ueno, H. Ago, R. R. Eady, T. Tosha, M. Yamamoto, S. S. Hasnain Science Advances, 7, eabd8523 (2021)); S. L. Rose. S. Baba, H. Okumura, S. V. Antonyuk, D. Sasakia, T. M. Hedison, M. Shanmugam, D. J. Heyes, N. S. Scrutton, T. Kumasaka,Takehiko Toshad , Robert R. Eadya , Masaki Yamamotod, and S. S. Hasnain. PNAS, 119 e2205664119 (2022); C. Gopalsingam and S. S. Hasnain, Curr Opin in Struc Biol 75:102420 (2022); |
| Description | Developmet of Spectroscopically validated MSOX movies of metalloenzymes |
| Organisation | Diamond Light Source |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Structural movies can be obtained by serially recording multiple structures (frames) from the same spot of a crystal using the MSOX serial crystallography approach. This method has been combined with on-line single crystal optical spectroscopy to capture the catalysis of CuNiRs from two Rhizobia species in action. We have provided the well diffracting crystals and all the know-how of enzyme mechanism. |
| Collaborator Contribution | An efficient on-line optical spectroscopic set up has been added to I24 with active participation of the beamline staff in our research where our team (including me) participated in on-site experiments. |
| Impact | A paper is to be submitted to Nature Structural Biology shortly |
| Start Year | 2022 |
| Description | Trangenic mice treatment plan established with Selenium based compounds |
| Organisation | Nagoya University |
| Country | Japan |
| Sector | Academic/University |
| PI Contribution | Novel Selenium-based compounds with therapeutic potential for SOD1-linked amyotrophic lateral sclerosis. |
| Collaborator Contribution | Validated protocols have been established to undertake transgenic experiments. These have shown clear disease onset delay of ebselen in transgenic ALS mice model holding encouraging promise for potential therapeutic compounds. We are expecting to extend this to our new award from MND association to non-covalent binders during a visit in May 2024 which will have an impact on "Development of non-covalent binders to SOD1 with scope for developing as PROTAC degraders of SOD1 for the treatment of MND". |
| Impact | Significant results have appeared in EBioMedicine 59 (2020) 102980 and another paper is under review with Nature Communications. |
| Start Year | 2020 |
| Description | UK-Japan symposium on Frontiers of Structural Biology, 2 DAYS Symposium (18-19 June 2024) at the University of Liverpool opened to the UK community with 26 invited and plenary speakers, 13 of which from Japan |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | The symposium celebrates the Japan-UK partnership and brings together wide range of experts from Japan covering all of the frontier structural biology approaches including cryo-electron microscopy (cryoEM), Synchrotron and Free Electron Laser X-ray crystallography and neutron crystallography. The symposium will have two full days of Lectures, the first day finishing with a Public lecture by the Nobel Laureate Richard Henderson and the second day with Lectures (open to public) from Sir Tom Blundell and Dr Harren Jhoti, FRS (President and Chief executive of Astex Pharmaceuticals). A PDF of the programme is attached. The symposium is supported by the University of Liverpool and RIKEN together with BBSRC, STFC and ILL. |
| Year(s) Of Engagement Activity | 2024 |
| URL | https://payments.liv.ac.uk/conferences-and-events/events-at-liverpool/institute-of-systems-molecular... |