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Elucidating the principles of the molecular organization of heteropolymeric tight junction strands

by Rahn, Hans-Peter , Fritzsche, Susanne , Walter, Maria , Gehring, Claudia , Wolburg, Hartwig , Turner, Jerrold R. , Blasig, Ingolf E. , Cording, Jimmi , Yu, Dan , Richter, Sandra , Hartwig, Jens , Piontek, Jörg

in Biochemistry / Biomedical and Life Sciences / Biomedicine / Blood-Brain Barrier - cytology / Blood-Brain Barrier - metabolism / brain / Cell Biology / Cells, Cultured / Claudins - chemistry / Claudins - genetics / Claudins - metabolism / coculture / electron microscopy / Energy transfer / fluorescence / HEK293 Cells / Humans / image analysis / Life Sciences / Molecular biology / Polymers / proteins / Research Article / tight junctions / Tight Junctions - chemistry / Tight Junctions - metabolism / Tissues

2011

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Elucidating the principles of the molecular organization of heteropolymeric tight junction strands

2011

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2011

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Journal Article

Elucidating the principles of the molecular organization of heteropolymeric tight junction strands

Rahn, Hans-Peter,

Fritzsche, Susanne,

Walter, Maria,

Gehring, Claudia,

Wolburg, Hartwig,

Turner, Jerrold R.,

Blasig, Ingolf E.,

Cording, Jimmi,

Yu, Dan,

Richter, Sandra,

Hartwig, Jens,

Piontek, Jörg

2011

Overview

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.

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Publisher

SP Birkhäuser Verlag Basel,Springer Nature B.V

Subject

Biochemistry

/ Biomedical and Life Sciences

/ Biomedicine

/ Blood-Brain Barrier - cytology

/ Blood-Brain Barrier - metabolism

/ brain

/ Cell Biology