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InstructThe latest webinar in the Instruct-ERIC Structure Meets Function series this month includes speakers who have accessed cutting-edge structural biology services through Instruct. The webinar series offers an insight into the very cutting edge of structural biology research, utilising the latest techniques available through Instruct-ERIC facilities and centres.
In May, we will hear from Heikki Saari of the University of Helsinki, whose Instruct R&D project helped to generate a Raman spectroscopy tool for extracellular vesicle analysis. We will also have a talk from Carine de Marcos Lousa of the University of Dundee, who accessed X-Ray and Cryo-EM technologies at the University of Leeds and the University of Oxford through Instruct.
The webinar will take place 20 May at 11:00 CEST - book your spot and register here.
Speaker: Heikki Saari, University of Helsinki
Title: Inline Raman spectroscopy for monitoring platelet extracellular vesicle purification with anion-exchange chromatography
Abstract: Extracellular vesicles (EVs) are relatively recently discovered biological nanoparticles that mediate intercellular communication. The development of new methods for the isolation and characterization of EVs is crucial to support further studies on these small and structurally heterogenous vesicles. Here, we demonstrate the use of an inline Raman detector in conjunction with anion exchange chromatography for the isolation of EVs from human platelets. Anion-exchange chromatography can be easily coupled with multiple inline detectors and provides an alternative to size-based methods for separating EVs from similar-sized impurities, such as lipoprotein particles. Our results show a notable separation of impurities from the EVs during anion-exchange chromatography, with Raman spectroscopy enabling us to identify functional groups beyond just bulk protein concentration in platelet EV samples. With this added information, it was possible to create a fingerprint for the EV- and impurity-fractions and to monitor any alterations in the purification process due to differences in the input sample. In conclusion, compared to conventional EV analysis methods, the inline Raman approach does not require additional hands-on work beyond the chromatographic EV purification and can provide detailed, real-time information about the sample and the purification process.
Speaker: Carine de Marcos Lousa, University of Dundee
Title: The Crystal Structure of Vacuolar Sorting Receptor
Abstract: Vacuolar sorting receptors (VSRs) are type I membrane proteins essential for plant development. Although their trafficking pathways have been extensively characterized, the molecular mechanisms underlying cargo recognition and release by their N-terminal lumenal domains remain poorly understood. We have elucidated the crystal structures of the lumenal region of Arabidopsis VSR1 in both cargo-free and cargo-bound states. These structures reveal the conformational changes associated with cargo binding and suggest a dynamic oligomerization mechanism that may regulate receptor function and trafficking. Our findings offer new insights into the molecular basis of VSR-mediated cargo sorting and propose a mechanistic model potentially applicable to other membrane trafficking receptors.
