Research

The HistoGenes project unites historians, archaeologists, geneticist, anthropologists, and specialists in bio-informatics, isotope analysis and other scientific methods in order to investigate human migration in the Carpathian Basin after the break down of the Roman Empire 400-900 CE. www.histogenes.org The Institute of Analytical Chemistry (IAC) at the University of Natural Resources and Life Sciences, Vienna will perform the analysis of strontium isotopic n(87Sr)/n(86Sr) ratios and multi-elemental patterns of teeth from individuals excavated at two Awar cemeteries in Austria. Furthermore, the IAC will cooperate with the other beneficiaries for interpretation of the results, which can only be done in an interdisciplinary team due to the complexity of the burial site. The gained information will allow to understand if individuals were local or migrated. This will advance our knowledge about population dynamics in a key period in European history.

Wider research context Of the five human heme peroxidases, evolved to perform widely divergent functions, thyroid peroxidase (TPO) is the least well studied. The multidomain membrane protein catalyses the biosynthesis of thyroid hormones, which are essential for metabolism, growth and development of the human body. With the use of hydrogen peroxide TPO catalyses the iodination and coupling of tyrosine residues on the surface of thyroglobulin (TG) in the thyroid gland. However, there are open questions about substrate selectivity, biochemical characteristics, the coupling mechanism and the roles of the individual domains. Critically, TPO is at the core of two autoimmune thyroid diseases that combined afflict nearly 150 million people but the only two clinically approved TPO inhibitors are not specific. Objectives This project will elucidate the biochemical and structural characteristics of TPO. The main goals are (I) understanding the kinetics and substrate specificity of TPO, (II) provide structural data of TPO alone and of relevant ligated states and (III) clarify the mode of action of TPO inhibitors. Approach In preliminary work an expression and purification protocol for truncated TPO variants was established. Importantly, it was found that it is possible to reconstitute and link the heme cofactor even after purification, yielding highly pure and enzymatically active recombinant TPO. This allows the first detailed spectroscopic, thermodynamic and structural study of the enzyme. This will include an analysis of the reaction kinetics of TPO using a range of methods, including stopped-flow UV-vis spectroscopy for pre-steady kinetics, analysis of the interaction with small molecules (i.e. inhibitors, ligands) and TG with thermodynamic and mass spectroscopic assays. Finally, this project aims to solve the X-ray crystal structure of TPO alone and in complex with biologically relevant substrates, ligands and inhibitors. Level of originality TPO is crucial in thyroid hormone biosynthesis. However, to date the available biochemical and structural data is scarce and does not allow (I) a clear rationalization of the structure-function relationship of the active site architecture and TPO reactivity, (II) the role of the additional TPO domains and (III) the design of new more specific inhibitors. This project aims to provide a complete enzymatic and structural characterization of TPO to address these issues. Primary researchers involved Vera Pfanzagl graduated from the international PhD program BioToP at BOKU in 2019. She worked primarily on structure-function relationships of heme enzymes and during her postdoc focused on human heme peroxidases. Within this Fellowship she aims to obtain her habilitation to advance her academic career. She will be supported and coached by Kristina Djinovic-Carugo, a highly recognized expert in structural biology and head of EMBLE Grenoble and Chris Oostenbrink (professor at BOKU), an expert in molecular modelling and simulation.

Project "Flame retardant treatment of wood using supercritical carbon dioxide" (FFG, Bridge-1 program, project number FO999903628) is a network project between BOKU, Kompetenzzentrum Wood KPlus and the companies NATEX (Austria) and Fritz Kohl GmbH & Co. KG. The impregnation of technical veneers with high-performance flame retardants using supercritical CO2 as a carrier medium is to be investigated with the aim of using these veneers in aircraft, automobile and shipbuilding. In the 3-year's project, R&D work is carried out in 4 central work packages (in addition to the APs Management and Dissemination). Based on a comprehensive literature study (WP 2) on P- and N-containing high-performance flame retardants, accompanying experiments investigating their dissolution and transport behavior in scCO2 as well as experiments targeting the introduction of functional groups for improved solubility or chemical fixation in wood, a selection of promising flame retardants for further work in WP 3-5 will be made. For these compounds, investigations into the solubility and transport behavior in scCO2 will be carried out in AP 3. This work allows the identification of particularly promising FSM and impregnation conditions (p, T, relaxation regime). Subsequently, tests are carried out to impregnate wood veneers, technical veneers and small solid wood samples in order to evaluate the loading success qualitatively and quantitatively. Accordingly, these tests are accompanied by comprehensive analysis, which serves to demonstrate the homogeneity of the loading, the degree of loading and the influence of the treatment on essential material properties. Data from these experiments can be used to draw important conclusions about improving process control. Finally, in WP 5, flame protection-relevant, mechanical, olfactory, morphological and colorimetric parameters will be collected for selected samples in order to finally evaluate the practical suitability of scCO2-supported impregnation of wood veneers and its transfer to solid wood. .