WWTF funded Project “Structure Zoom” (LS17-008)

Zooming in on protein functional sites with atomic resolution – an integrated chemistry approach for structural biology.

In the Structure Zoom project, we aim to apply chemical protein synthesis techniques to generate site-specifically labeled and modified proteins for structural biology of key proteins in neurodegeneration and muscle disease. Our unique interdisciplinary expertise in protein synthesis, site-specific modification and structural biology makes it feasible to design and synthesize protein constructs that provide precise information about structural features and dynamics by NMR and small angle scattering (SAS). The innovative aspects of the project lie in our ability to generate specifically isotope-labeled and/or posttranslationally modified proteins by combining synthesis of building blocks with protein synthesis. With this versatile approach, we can access protein samples that have so far not been available for NMR and SAS studies. As a proof-of-principle, we will apply our chemical toolbox of labeling and semisynthesis strategies to two proteins (tau-4 and FATZ) and use the resulting structural and dynamic data on the interactions of these proteins with their respective binding partners to provide insights into their molecular structural organization. Furthermore, this project establishes a new partnership between the peptide and protein chemistry group at the University of Vienna and the structural biology center at the Max F. Perutz Laboratories, enabling unique interdisciplinary training of young researchers at the interface of these two disciplines.

Participants:

Institute of Biological Chemistry, Faculty of Chemistry, University of Vienna

Christian F. W. Becker (Coordinator)

School of Biomedical Sciences, University of Queensland

Anne C. Conibear

Max F Perutz Laboratories, University of Vienna

Kristina Djinovic-Carugo, Robert Konrat

Institute of Organic Chemistry, Faculty of Chemistry, University of Vienna

Roman Lichtenecker

Publications:

Conibear, A.C., Watson, E.E., Payne, R.J., C., Becker, C.F. (2018) Native chemical ligation in protein synthesis and semi-synthesis. Chem. Soc. Rev., 47, 9046-9068.

Conibear. A.C., Rosengren, K.J., Becker, C.F., Kaehlig, H. (2019) Random coil shifts of posttranslationally modified amino acids. J. Biomol. NMR, 73, 587-599.

Ellmer, D., Brehs, M., Haj‐Yahya, M., Lashuel, H.A., Becker, C.F. (2019) Single Posttranslational Modifications in the Central Repeat Domains of Tau4 Impact its Aggregation and Tubulin Binding. Angew. Chem. Int. Ed., 58, 1616–1620.

Mlynek, G., Kostan, J., Leeb, S., and Djinović-Carugo, K. (2020) Tailored Suits Fit Better: Customized Protein Crystallization Screens. Crystal Growth & Design 20, 984-994

Pinotsis, N., Zielinska, K., Babuta, M., Arolas, J. L., Kostan, J., Khan, M. B., Schreiner, C., Salmazo, A., Ciccarelli, L., Puchinger, M., Gkougkoulia, E. A., Ribeiro, E. A., Jr., Marlovits, T. C., Bhattacharya, A., and Djinović-Carugo, K. (2020) Calcium modulates the domain flexibility and function of an alpha-actinin similar to the ancestral alpha-actinin. Proc Natl Acad Sci U S A

Hackl, S., Ng, X.W., Lu, D., Wohland, T., Becker, C.F.W. (2021) Cytoskeleton-dependent clustering of membrane-bound prion protein on the cell surface. J. Biol. Chem.

Niederacher, G., Urwin, D., Tremethick, D.J., Rosengren, K.J., Becker, C.F.W., Conibear, A.C. (2021) Site-specific modification and segmental isotope labelling of HMGN1 reveals long-range conformational perturbations caused by posttranslational modifications. RCS Chem. Biol.

Kostan, J., Pavšič, M., Puž, V., Schwarz, T. C., Drepper, F., Molt, S., Graewert, M. A., Schreiner, C., Sajko, S., Van Der Ven, P. F. M., Svergun, D. I., Warscheid, B., Konrat, R., Fürst, D. O., Lenarčič, B., and Djinović-Carugo, K. (2021) Molecular basis of F-actin regulation and sarco-mere assembly via myotilin. PLOS Biol.

Sponga, A., Arolas, J. L., Schwarz, T. C., Jeffries, C. M., Chamorro, A. R., Kostan, J., Ghisleni, A., Drepper, F., Polyansky, A., Ribeiro, E. D. A., Pedron, M., Zawadzka-Kazimierczuk, A., Mlynek, G., Peterbauer, T., Doto, P., Schreiner, C., Hollerl, E., Mateos, B., Geist, L., Faulkner, G., Kozminski, W., Svergun, D. J., Warscheid, B., Zagrovic, B., Gautel, M., Konrat, R., and Djinović-Carugo, K. (2021) Order from disorder in the sarcomere: FATZ forms a fuzzy but tight complex and phase-separated condensates with {alpha}-actinin. In press in Sci Adv