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      Flow Alignment of Extracellular Vesicles: Structure and Orientation of Membrane-Associated Bio-macromolecules Studied with Polarized Light.

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          Abstract

          Extracellular vesicles (EVs) are currently in scientific focus, as they have great potential to revolutionize the diagnosis and therapy of various diseases. However, numerous aspects of these species are still poorly understood, and thus, additional insight into their molecular-level properties, membrane-protein interactions, and membrane rigidity is still needed. We here demonstrate the use of red-blood-cell-derived EVs (REVs) that polarized light spectroscopy techniques, linear and circular dichroism, can provide molecular-level structural information on these systems. Flow-linear dichroism (flow-LD) measurements show that EVs can be oriented by shear force and indicate that hemoglobin molecules are associated to the lipid bilayer in freshly released REVs. During storage, this interaction ceases; this is coupled to major protein conformational changes relative to the initial state. Further on, the degree of orientation gives insight into vesicle rigidity, which decreases in time parallel to changes in protein conformation. Overall, we propose that both linear dichroism and circular dichroism spectroscopy can provide simple, rapid, yet efficient ways to track changes in the membrane-protein interactions of EV components at the molecular level, which may also give insight into processes occurring during vesiculation.

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          Author and article information

          Journal
          Chembiochem
          Chembiochem : a European journal of chemical biology
          Wiley
          1439-7633
          1439-4227
          Mar 16 2018
          : 19
          : 6
          Affiliations
          [1 ] Institute of Materials and Environmental Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, P. O. Box 286, 1519, Budapest, Hungary.
          [2 ] Department of Biology and Biological Engineering, Chalmers University of Technology, Chemical Biology, Kemigården 4, 41296, Göteborg, Sweden.
          [3 ] Department of Biophysics and Radiation Biology, Semmelweis University, Tűzoltó u. 37-47, 1094, Budapest, Hungary.
          [4 ] Department of Chemistry and Chemical Engineering, Chemistry and Biochemistry, Chalmers University of Technology, Kemigården 4, 41296, Göteborg, Sweden.
          Article
          10.1002/cbic.201700378
          29237098
          82ab541a-d4ee-4fe5-a538-9649e0eec140
          History

          liposomes,polarized light spectroscopy,probe molecules,transition moment orientation,extracellular vesicles

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