News

Our Manuscript published in ChemCommun

Our Manuscript published in ChemCommun

The synthesis of specifically isotope labelled Fluorotryptophan and its use in mammalian cell-based protein expression for 19F-NMR applications.

Giorgia Toscano,   Martina Rosati,   Letizia Barbieri,   Katharina Maier,   Lucia Banci,   Enrico Luchinat,   Robert Konrat  and  Roman Josef Lichtenecker  

Chem. Commun., 2024, Accepted Manuscript

19F nuclei serve as versatile sensors for detecting protein interactions and dynamics in biomolecular NMR spectroscopy. Although various methods have been developed to incorporate fluorine-containing aromatic residues into proteins using E. coli or cell-free expression techniques, similar approaches for protein production in mammalian cell lines remain limited. Here, we present a cost-effective synthetic route to obtain selectively deuterated, carbon-13 labeled fluorotryptophan and demonstrate its use in introducing 19F-13C spin pairs into Carbonic Anhydrase 2 and Superoxide Dismutase, following an expression protocol utilizing HEK cells.

Our new paper is online

Our new paper is online

Synthesis of a 13C/2H Labeled Building Block to Probe the Phosphotyrosine Interactome Using Biomolecular NMR Spectroscopy

Sarah Kratzwald, Thomas C. Schwarz, Karin Ledolter, Matus Hlavac, Manuel Felkl, Christian F. W. Becker, Robert Konrat and Roman J. Lichtenecker

ChemBioChem 2024 doi.org/10.1002/cbic.202400663

A protected 13C/2H-labeled phosphotyrosine (pTyr) building block was synthesized from commercially available isotope sources for use in solid-phase peptide synthesis (SPPS). The resulting isolated ϵ-aromatic 13C-1H correlations provide valuable data for studying interactions with pTyr-binding proteins at atomic resolution using NMR spectroscopy techniques. As an initial demonstration, intermolecular NOE and chemical shift perturbation data were recorded to visualize the binding of a pTyr-containing peptide, derived from the platelet-derived growth factor to the PLCγ-1 SH2 domain.

DoSChem awardees

DoSChem awardees

Giorgia Toscano (Lichtenecker group) and Alexandra Ptaszek (Konrat group) are the new awardees in the "New Ideas" program of the Doctoral" School in Chemistry of our Faculty. Their project Automating Methyl Assignment in Protein NMR by Machine Learning Chemical Shift Predictions and Isotopic Labelling will be funded within the DoSChem with a duration of three months. Gongratulations Giorgia and Alexandra and all the best for your project! 

New Team Member

New Team Member

Cora Döge started as a new bachelor student in the group. In her project, she mainly focuses on synthetic routes to decorate isotope labeled amino acids with protection groups. The resulting molecules will be used as building blocks in solid phase peptide synthesis. Welcome!

Paper alert

Characterization of conformational states of the homodimeric enzyme fluoroacetate dehalogenase by 19F-13C two-dimensional NMR

This paper by Scott Prosser and his team at the University of Toronto describes the NMR study of the 13C6 fluorotryptophan labeled enzyme fluoroacetate dehalogenase, which was overexpressed in E. coli using isotope labeled 5-fluoroanthranilic acid as a fTrp precursor.

The manuscript is online at: RSC Chem Bio doi: 10.1039/d4cb00176a

Master defense

Oct. 18th 2024: Noel Riedl successfully defended his Master Thesis of a project he conducted at the company Valdospan! Congratulations!

Cycling at the Neusiedler See

Cycling at the Neusiedler See

Members of our group met with people from Mag-Lab and the AG Konrat on August 24th 2024 to cycle round the Neusiedler See. 

We met at Neusiedl am See, where we headed south to Illmitz and took the ferry to Mörbisch. From there, we passed Rust and Purbach to reach our starting point again. The hot temperature could not stop us from enjoying a wonderful day in Burgenland. Thank you for joining. 

Advancing Cost-Effective Side-Chain Isotope Labeling for Proteins Expressed in Human Cells

Advancing Cost-Effective Side-Chain Isotope Labeling for Proteins Expressed in Human Cells

Side-chain isotope labeling is an essential technique for studying protein structure, dynamics, and interactions through NMR spectroscopy. While labeling procedures are now standard for E. coli and cell-free expression systems, proteins produced in mammalian cells typically require costly isotope-labeled amino acids to introduce carbon-13 and deuterium. In a collaborative research project between CERM-University of Florence, the University of Vienna, and Mag-Lab, we demonstrated that using cost-effective alpha-ketoacid precursors in mammalian cell culture media enables selective labeling without isotope scrambling. These labeled proteins are then directly used to study conformational changes upon ligand binding through in-cell and in-lysate NMR spectroscopy.

Read the full story in an open access article of the Journal of Biomolecular NMR:

https://link.springer.com/article/10.1007/s10858-024-00447-6

Protein NMR, isotope labeling and chemical shift calculations form the basis for a rapid and efficient structure optimization process of initial low-affinity binders

A recent research article published in the Journal of Medicinal Chemistry presents our latest findings from a collaborative project involving the University of Vienna, Boehringer-Ingelheim, and Mag-Lab, with financial support from the Christian Doppler Society. Our innovative method employs protein NMR spectroscopy, combined with aliphatic residue methyl labeling to directly measure CH-hydrogen bonding interactions. The resulting data on ligand-protein complex geometries helps to understand the residual mobility of ligands within their binding pockets, indirectly reflecting binding energies. These insights enable medicinal chemists to make well-informed decisions during accelerated structure optimization processes, even in the absence of protein-ligand cocrystal structures.

Read the full story in the open access article: https://pubs.acs.org/doi/10.1021/acs.jmedchem.4c01128

Retreat with the Groups of Paul Schanda and Frans Mulder

We spent very interesting three days starting from June 19th 2024 in Winterbach near the Ötscher to discuss the possibilities of biomolecular NMR and isotope labeling. Great fun and great science in a wonderful landscape.

5th OC SYMPOSIUM for Students

5th OC SYMPOSIUM for Students

The OC Symposium for students took place in the Auer-von-Welsbach HS on June 19th 2024.

Several students presented their work and Walther Schmid memorial prizes were awarded to two students for their outstanding bachelor theses. In addition to that, the Institute celebrated Harry Martin on the occasion of his retirement. From our group, Sarah gave a lecture talking about her endaveours within the scope of her Master's project.

The International Workshop on Advanced Isotopic Labelling Methods in Grenoble 2024

The International Workshop on Advanced Isotopic Labelling Methods in Grenoble 2024

The AILM 2024 took place at the EPN campus of Grenoble from May 28th to May 31st with Darja and Giorgia presenting their PhD projects in oral lecture contributions. As always, the conference was a wonderful opportunity to discuss the recent developments in the field of isotope labelling and its application to the study of biomolecular structure and dynamics.

41st Vienna City Marathon

41st Vienna City Marathon

Two teams combining the running power of the University labs and Mag-Lab participated in the relay competition of the Vienna City Marathon. A wonderful event and a great experience with a lot of fun thanks to Sarah, Alisa, Giorgia, Ola, Sven, Salvatore, Roman and Robert and of course all our fans and supporters.

DosChem Retreat 2024 in Röjtökmuzsaj

Together with the Böttcher Group we organized the DosChem retreat 2024 in Szedonia Castle (13-15.3.2024; Hungary). We enjoyed three days of science with 11 student talks, 36 posters presentations and our guest speakers with highly interesting contributions from their corresponding research fields. Darja Rohden was awarded the best student talk by the participating students and PIs. Congratulations!

Selective Methine Labeling to Probe Protein-Ligand Interactions

Precise information regarding the interaction between proteins and ligands at molecular resolution is crucial for effectively guiding the optimization process from initial hits to lead compounds in early stages of drug development. Giorgia Toscano introduced a novel aliphatic side chain isotope-labeling scheme to directly probe interactions between ligands and aliphatic sidechains using NMR techniques. Together with Theresa Höfurthner, Robert Konrat and our collaborators of the Christian Doppler Lab for High Content Structural Biology and Biotechnology, we demonstrated the applicability of this method by analyzing 1H chemical shift perturbation resulting from CH-π interaction of Hβ-Val and Hγ-Leu as CH donors with corresponding ligand aromatic moieties as π acceptors. Our studies were published in ChemBioChem on Jan 31st 2024.

Team News

Alisa Wimmer joined our Team on September 1st 2023 in the Organic Synthesis Lab to do her Master's project. Alisa is working in the company Mag-Lab, where she is an important factor to push the frontiers of isotope labeling further.

Welcome Alisa!

UniVie Connect: Academic and Pharma Research

UniVie Connect: Academic and Pharma Research

We participated in the UniVie Connect: Academic and Pharma Research event on June 30th 2023 and learned a lot about companies, funding opportunities and University-industry cooperations. In addition to that, we met company representatives and other academic researchers in interesting round table discussions.

Faculty football tournament

Faculty football tournament

Football tournament on the 26th of June at the DSV Fortuna pitch! Fantastic football with wonderful organization but one of these days when some people have to realize that they're not 20 anymore...

Team News

Katharina Maier joined the team for her bachelor thesis in May 2023. She works on new methods to install 13C-fluorine combinations in target proteins.

Inaugural Biochemistry Symposium

Inaugural Biochemistry Symposium

We visited the new Instutute of Biochemistry at JKU Linz (May 23rd and 24th) to celebrate the Inaugural Biochemistry Symposium and its head Prof. Frans Mulder. Fifteen renowned international scientists presented their work on Biomolecular structure and dynamics. A great event with many interesting discussions!

Doctoral School in Chemistry

Doctoral School in Chemistry

The group attended the DosChem retreat 2023 in Fürstenfeld March 22-24. Giorgia introduced her work in a talk and Darja contributed a poster presentation. With Clemens Decristoforo (Uni Innsbruck), Pablo Rivera Fuentes (Uni Zürich) and Paolo Melchiorre (Uni Bologna) we enjoyed three guest speakers in addition to all the great contributions from our PhD students.

NMR Symposium

Robert Konrat's 60th - From Dynamics to Disorder and Beyond

From the 28th to the 29th of September 2022, the Biomolecular NMR community celebrated the 60th birthday of Robert Konrat with several talks by Robert's former students and collaboration partners. The event also provided a stimulating environment for younger scientists to present their work and consequently Darja, Giorgia and Sarah showed their recent results on scientific posters.

Conference participation

Österreichische Chemietage

Our group participated in the Conference of the Austrian Chemical Society at the Technical University of Vienna.

Conference participation

Student Symposium of the Vienna Chemistry Doctoral School

The PhD students of our faculty showed their recent work at the 2nd DosChem International Student Symposium. Giorgia and Darja presented posters and gave insights into their projects in vivid discussions with their peer students.

Team News

Cherilyn Cikan

did a great job and defended her Master project entitled "Synthesis of stable heavy isotope containing small molecules" on the 05.09.2022. Congratulations!

Team News

Darja Rohden

joined our team in July 2022. Darja obtained her masters degree in the group of Prof. Davide Bonifazi and is now part of the Allospace program. Within the scope of her PhD project, Darja will elucidate the allosteric network of target proteins on an atomic resolution. She will combine Organic Chemistry with solid state protein NMR spectroscopy, which will be performed in the group of Paul Schanda at the ISTA.

Teaching

We returned to the lecture halls

After a long time of Covid restrictions and many online courses, we are back at the lecture halls with four courses in SS2022:

270032 VO Organic Chemistry for Highschool Teachers - -Chemistry

270055 SE Actual Problems in Bioorganic Chemistry

270071 VO Introduction into carbohydrates - Introduction into carbohydrate chemistry and biological chemistry

270084 VO Selected topics in bioorganic chemistry