ERC FUNDED PROJECTS

Crucial processes within cells depend on specific non-covalent interactions which mediate the assembly of proteins and other biomolecules. Deriving structural information to understand the function of these complex systems is the primary goal of Structural Biology. In this application, the recently developed LILBID method (Laser Induced Liquid Bead Ion Desorption) will be optimized for investigation of macromolecular complexes with a mass accuracy two orders of magnitude better than in 1st generation spectrometers. Controlled disassembly of the multiprotein complexes in the mass spectrometric analysis while keeping the 3D structure intact, will allow for the determination of complex stoichiometry and connectivity of the constituting proteins. Methods for such controlled disassembly will be developed in two separate units of the proposed LILBID spectrometer, in a collision chamber and in a laser dissociation chamber, enabling gas phase dissociation of protein complexes and removal of excess water/buffer molecules. As a third unit, a chamber allowing determination of ion mobility (IM) will be integrated to determine collisional cross sections (CCS). From CCS, unique information regarding the spatial arrangement of proteins in complexes or subcomplexes will then be obtainable from LILBID. The proposed design of the new spectrometer will offer fundamentally new possibilities for the investigation of non-covalent RNA, soluble and membrane protein complexes, as well as broadening the applicability of non-covalent MS towards supercomplexes.

1 264 477 €

Start date: 2014-02-01, End date: 2019-01-31

The development of longer lasting, higher energy density and cheaper rechargeable batteries represents one of the major technological challenges of our society, batteries representing the limiting components in the shift from gasoline-powered to electric vehicles. They are also required to enable the use of more (typically intermittent) renewable energy, to balance demand with generation. This proposal seeks to develop and apply new NMR metrologies to determine the structure and dynamics of the multiple electrode-electrolyte interfaces and interphases that are present in these batteries, and how they evolve during battery cycling. New dynamic nuclear polarization (DNP) techniques will be exploited to extract structural information about the interface between the battery electrode and the passivating layers that grow on the electrode materials (the solid electrolyte interphase, SEI) and that are inherent to the stability of the batteries. The role of the SEI (and ceramic interfaces) in controlling lithium metal dendrite growth will be determined in liquid based and all solid state batteries. New DNP approaches will be developed that are compatible with the heterogeneous and reactive species that are present in conventional, all-solid state, Li-air and redox flow batteries. Method development will run in parallel with the use of DNP approaches to determine the structures of the various battery interfaces and interphases, testing the stability of conventional biradicals in these harsh oxidizing and reducing conditions, modifying the experimental approaches where appropriate. The final result will be a significantly improved understanding of the structures of these phases and how they evolve on cycling, coupled with strategies for designing improved SEI structures. The nature of the interface between a lithium metal dendrite and ceramic composite will be determined, providing much needed insight into how these (unwanted) dendrites grow in all solid state batteries. DNP approaches coupled with electron spin resonance will be use, where possible in situ, to determine the reaction mechanisms of organic molecules such as quinones in organic-based redox flow batteries in order to help prevent degradation of the electrochemically active species. This proposal involves NMR method development specifically designed to explore a variety of battery chemistries. Thus, this proposal is interdisciplinary, containing both a strong emphasis on materials characterization, electrochemistry and electronic structures of materials, interfaces and nanoparticles, and on analytical and physical chemistry. Some of the methodology will be applicable to other materials and systems including (for example) other electrochemical technologies such as fuel cells and solar fuels and the study of catalysts (to probe surface structure).

3 498 219 €

Start date: 2019-10-01, End date: 2024-09-30

Many significant global challenges in catalysis for energy and sustainable chemistry have already been solved in nature. Metalloenzymes within microorganisms catalyse the transformation of carbon dioxide into simple carbon building blocks or fuels, the reduction of dinitrogen to ammonia under ambient conditions and the production and utilisation of dihydrogen. Catalytic sites for these reactions are necessarily based on metals that are abundant in the environment, including iron, nickel and molybdenum. However, attempts to generate biomimetic catalysts have largely failed to reproduce the high activity, stability and selectivity of enzymes. Proton and electron transfer and substrate binding are all finely choreographed, and we do not yet understand how this is achieved. This project develops a suite of new experimental infrared (IR) spectroscopy tools to probe and understand mechanisms of redox metalloenzymes in situ during electrochemically-controlled steady state turnover, and during electron-transfer-triggered transient studies. The ability of IR spectroscopy to report on the nature and strength of chemical bonds makes it ideally suited to follow the activation and transformation of small molecule reactants at metalloenzyme catalytic sites, binding of inhibitors, and protonation of specific sites. By extending to the far-IR, or introducing mid-IR-active probe amino acids, redox and structural changes in biological electron relay chains also become accessible. Taking as models the enzymes nitrogenase, hydrogenase, carbon monoxide dehydrogenase and formate dehydrogenase, the project sets out to establish a unified understanding of central concepts in small molecule activation in biology. It will reveal precise ways in which chemical events are coordinated inside complex multicentre metalloenzymes, propelling a new generation of bio-inspired catalysts and uncovering new chemistry of enzymes.

1 997 286 €

Start date: 2019-03-01, End date: 2024-02-29

The demand for donated organs vastly outnumbers the supply, leading each year to the death of thousands of people and the suffering of millions more. Engineered tissues and organs following Tissue Engineering approaches are a possible solution to this problem. However, a prevascularization solution to irrigate complex engineered tissues and assure their survival after transplantation is currently elusive. In the human body, complex organs and tissues irrigation is achieved by a network of blood vessels termed capillary bed which suggests such a structure is needed in engineered tissues. Previous approaches to engineer capillary beds reached different levels of success but none yielded a fully functional one due to the inability in simultaneously addressing key elements such as correct angiogenic cell populations, a suitable matrix and dynamic conditions that mimic blood flow. CapBed aims at proposing a new technology to fabricate in vitro capillary beds that include a vascular axis that can be anastomosed with a patient circulation. Such capillary beds could be used as prime tools to prevascularize in vitro engineered tissues and provide fast perfusion of those after transplantation to a patient. Cutting edge techniques will be for the first time integrated in a disruptive approach to address the requirements listed above. Angiogenic cell sheets of human Adipose-derived Stromal Vascular fraction cells will provide the cell populations that integrate the capillaries and manage its intricate formation, as well as the collagen required to build the matrix that will hold the capillary beds. Innovative fabrication technologies such as 3D printing and laser photoablation will be used for the fabrication of the micropatterned matrix that will allow fluid flow through microfluidics. The resulting functional capillary beds can be used with virtually every tissue engineering strategy rendering the proposed strategy with massive economical, scientific and medical potential

1 499 940 €

Start date: 2018-11-01, End date: 2023-10-31