Elucidating the principles of the molecular organization of heteropolymeric tight junction strands
| Autor: | Dan Yu, Ingolf E. Blasig, Maria Walter, Hartwig Wolburg, Jerrold R. Turner, Claudia Gehring, Susanne Fritzsche, Jimmi Cording, Hans-Peter Rahn, Jens Hartwig, Sandra Richter, Jörg Piontek |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Biology
Blood–brain barrier Article Tight Junctions law.invention Protein–protein interaction 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine law medicine Humans Claudin Molecular Biology Cells Cultured Barrier function 030304 developmental biology Pharmacology 0303 health sciences Tight junction Cell Biology Claudins FRET Cell biology HEK293 Cells Förster resonance energy transfer medicine.anatomical_structure Blood-Brain Barrier Paracellular transport Molecular Medicine Electron microscope 030217 neurology & neurosurgery |
| Zdroj: | Cellular and Molecular Life Sciences; Vol 68 Cellular and molecular life sciences, 68(23): 3903–3918 |
| ISSN: | 1420-682X |
| DOI: | 10.1007/S00018-011-0680-z |
| Popis: | Paracellular barrier properties of tissues are mainly determined by the composition of claudin heteropolymers. To analyze the molecular organization of tight junctions (TJ), we investigated the ability of claudins (Cld) to form homo- and heteromers. Cld1, -2, -3, -5, and -12 expressed in cerebral barriers were investigated. TJ-strands were reconstituted by claudin-transfection of HEK293-cells. cis-Interactions and/or spatial proximity were analyzed by fluorescence resonance energy transfer inside and outside of strands and ranked: Cld5/Cld5 > Cld5/Cld1 > Cld3/Cld1 > Cld3/Cld3 > Cld3/Cld5, no Cld3/Cld2. Classic Cld1, -3, and -5 but not non-classic Cld12 showed homophilic trans-interaction. Freeze-fracture electron microscopy revealed that, in contrast to classic claudins, YFP-tagged Cld12 does not form homopolymers. Heterophilic trans-interactions were analyzed in cocultures of differently monotransfected cells. trans-Interaction of Cld3/Cld5 was less pronounced than that of Cld3/Cld1, Cld5/Cld1, Cld5/Cld5 or Cld3/Cld3. The barrier function of reconstituted TJ-strands was demonstrated by a novel imaging assay. A model of the molecular organization of TJ was generated. |
| Databáze: | OpenAIRE |
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