InstructThe latest webinar in the Instruct-ERIC Structure Meets Function series will be hosted by Instruct Netherlands on 12th April 2022, 11:00 - 12:30 CET. Register for the webinar here.
Featuring expert speakers from Instruct Centres across Europe, Instruct-ERIC Webinar Series: Structure Meets Function highlights some of the latest developments in structural biology, demonstrating how integrative methods are enabling scientists to decipher the mechanisms that underpin health and disease.
Watch the previous webinars in the series here.
Moderator:
Talk 1: A Real-Time Approach to Study Cellular Metabolism in Hepatocytes
Speaker: Simone Fjordside
Abstract: An In-Cell NMR Bioreactor allows for real-time metabolomic studies in vitro. Here we intend to establish a robust NMR Bioreactor setup for the study of mammalian liver cells that are encapsulated in a hydrogel with continuous supply of fresh medium. With use of the bioreactor NMR spectra were successfully recorded continuously on living and metabolically active hepatocytes. Due consideration is given to the improvement of cellular viability.
Talk 2: Reconstituting the 2D world of signalling proteins – exploring the role of membrane topography for information transfer
Speaker: Kristina Ganzinger, AMOLF
Abstract: Cellular signalling - the transmission and processing of information by cells - is an essential feature of life and critical to coordinating the immune response. Cytokines – a family of small signalling proteins – play a pivotal role in orchestrating both innate and adaptive immune responses. The cytokines interleukin-2 (IL-2) and interleukin-15 (IL-15), for example, set T- and NK-cells on course to respond to pathogens and tumours. IL-2 and IL-15 bind to a shared heterodimeric receptor consisting of IL-2/15Rβ and IL-2/15Rγ in complex with a private subunit. Signalling is transduced solely through the shared receptors, with no apparent structural differences between the IL-2 and IL-15 quaternary complexes. However, distinct and competing functional differences in IL-2 and IL-15 signalling indicate the existence of a ligand discrimination mechanism. To elucidate potential ligand discrimination mechanisms based on the kinetics of receptor assembly, we apply multi-colour single-molecule imaging on a reconstituted membrane-receptor system to follow individual receptor subunits and their assembly upon ligand binding. I will present on our first results and technical developments on the way, from building a bespoke four-colour single-molecule total internal reflection fluorescence (TIRF) microscope to developing a new single-molecule tracking tool based on DNA-PAINT.
Talk 3: Structure of a proteo-lipidic viral tail-tube for viral genome delivery
Speaker: Nicola Abrescia, CIC bioGUNE
Abstract: We have been involved in studying the lipid-containing icosahedral bacteriophage PRD1 as a model system for membrane-containing viruses by cryo-EM and X-ray techniques. In 2010, we become interested in the mechanism of genome translocation across the cell envelope. By combining SPA cryo-EM and cryo-electron tomography (cryo-ET) we showed that to deliver its double stranded DNA genome, the icosahedral protein-rich virus membrane transforms into a tubular structure protruding from one of the twelve vertices of the capsid (1-3). The internal viral membrane remodels and self-assembles triggered by changes in osmolarity and loss of capsid-membrane interactions. These events are a consequence of the de-capping of the vertices upon spike proteins interaction with cell surface receptors (as today still unknown). From earlier studies the tail tube appears to be structured by membrane-associated proteins (1, 4).
Since 2019 we have been interested in resolving the structure and the principles ruling the self-assembly of this proteo-lipidic DNA shuttling viral tube.
