Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
SHR
Neuro
Krebs
Kardio
Stoffw
Microb
Lipid
Lösung des offenen Rätsels von TRPC3 durch Photopharmakologie
- Abstract
- • Wider research context / theoretical framework: TRPC3 is a distinct member of TRPC family. This channel is expressed in excitable and non-excitable cells. By non-selectively conducting cations even at its resting state, TRPC3 promotes cellular processes in both health and disease. However, there are only a few modulators of this channel available. To precisely design new therapeutic ligands, it is of high importance to gain knowledge about TRPC3 protein structure and, consequently, its gating mechanism. Recently, two TRPC3 structures were revealed by cryo electron microscopy (cryo-EM). Nonetheless, neither of them captured open pore architecture. Hence, the open conformations is the critical missing piece of information to solve the TRPC3 gating puzzle. This knowledge is essential for comprehension of the molecular function of TRPC3 and definition of possible ligand binding sites to control this channel is health and, especially, disease.
• Hypotheses / research questions / objectives: Here, we propose to succeed in capturing open pore structures of TRPC3 by stabilizing and synchronizing the channels at a high open probability using photochromic ligands and in addition by preserving their lipid environment during the protein preparation for cryo-EM.
• Approach / methods: We will screen two expression systems for suitability to maintain TRPC3 in open conformation. We will utilize photopharmacological tools for a temporal control over TRPC3 gating during protein extraction and cryo-EM. Structural details obtained from cryo-EM will be followed by structure-guided mutagenesis and evaluated in a multifunctional live cell approach combining Ca2+ imaging and electrophysiology.
• Level of originality / innovation: With our project, we will introduce a novel approach to control protein conformations during cryo-EM by photoswitchable ligands. Our work will unravel TRPC3 open architecture and promote the development of new concepts for therapeutic targeting of these channels.
• Primary researchers involved: The research team will include profound experts in TRPC3 electrophysiology and Ca2+ signaling Dr. Oleksandra Tiapko and Dr. Klaus Groschner (Medical University of Graz, Austria), experts in membrane lipids Dr. Anita Emmerstorfer-Augustin (Graz University of Technology) and in structural biology and electron microscopy Dr. Christine Ziegler (University of Regensburg).
- Schlagworte
- Ca2+ kanal
- Cryo-EM
- Ionenkanal
- Strukturbiologie
- TRPC3
- Projektleitung:
- Tiapko Oleksandra
- Laufzeit:
- 01.01.2022-31.10.2025
- Programm:
- FWF Einzelprojekt
- Art der Forschung
- Angewandte Forschung
- Mitarbeiter*innen
- Tiapko O., Projektleiter*in
- Baron J., Projektmitarbeiter*in
- Beteiligte MUG-Organisationseinheiten
- Lehrstuhl für Medizinische Physik und Biophysik
- Projektpartner
- Graz University of Technology, Österreich
- University of Graz, Österreich
- University of Regensburg, Deutschland