Human Claudin-7 cis-Interactions Are Not Crucial for Membrane-Membrane (Trans-) Interactions

Publication type:
Journal article
Metadata:
Abstract
<jats:p>Human Claudin-7 (Cldn7) is a member of the Claudin (Cldn) superfamily. <jats:italic>In vivo</jats:italic>, these proteins form tight junctions, which establish constricted connections between cells. Cldns oligomerize within the membrane plane (= <jats:italic>cis</jats:italic>-interaction), and also interact with Cldns from adjacent cells (= <jats:italic>trans</jats:italic>-interaction). Interactions of Cldns are typically studied <jats:italic>in vivo</jats:italic> and structural analyses of isolated Cldns are limited. Here, we describe heterologous expression in <jats:italic>E. coli</jats:italic> and purification of human Cldn7, enabling <jats:italic>in vitro</jats:italic> analyses of the isolated protein using detergent and model membrane systems. Cldn7 exists as a monomer, hexamer, and various higher oligomers in micelles. While only limited unfolding of the protein was observed in the presence of the anionic detergent sodium dodecyl sulfate, decreased ionic strength did affect Cldn7 <jats:italic>cis</jats:italic>-interactions. Furthermore, we identified two amino acids which mediate electrostatic <jats:italic>cis</jats:italic>-interactions and analyzed the impact of disturbed <jats:italic>cis</jats:italic>-interaction on <jats:italic>trans</jats:italic>-contacts via atomic force microscopy and monitoring Förster resonance energy transfer between fluorescently labeled Cldn7-containing proteoliposomes. Our results indicate that Cldn7 <jats:italic>cis</jats:italic>-oligomerization might not be a prerequisite for establishing <jats:italic>trans</jats:italic>-contacts.</jats:p>
Autoren
  • Lena Ahlswede
  • Carmen Siebenaller
  • Benedikt Junglas
  • Nadja Hellmann
  • Dirk Schneider
DOI
10.3389/fmolb.2022.908383
eISSN
2296-889X
Zeitschrift
Frontiers in Molecular Biosciences
Online publication date
2022
Status
Published online
Herausgeber
Frontiers Media SA
Herausgeber URL
http://dx.doi.org/10.3389/fmolb.2022.908383
Datum der Datenerfassung
2022
Titel
Human Claudin-7 cis-Interactions Are Not Crucial for Membrane-Membrane (Trans-) Interactions
Ausgabe der Zeitschrift
9

Data source: Crossref

Other metadata sources:
Abstract
Human Claudin-7 (Cldn7) is a member of the Claudin (Cldn) superfamily. <i>In vivo</i>, these proteins form tight junctions, which establish constricted connections between cells. Cldns oligomerize within the membrane plane (= <i>cis</i>-interaction), and also interact with Cldns from adjacent cells (= <i>trans</i>-interaction). Interactions of Cldns are typically studied <i>in vivo</i> and structural analyses of isolated Cldns are limited. Here, we describe heterologous expression in <i>E. coli</i> and purification of human Cldn7, enabling <i>in vitro</i> analyses of the isolated protein using detergent and model membrane systems. Cldn7 exists as a monomer, hexamer, and various higher oligomers in micelles. While only limited unfolding of the protein was observed in the presence of the anionic detergent sodium dodecyl sulfate, decreased ionic strength did affect Cldn7 <i>cis</i>-interactions. Furthermore, we identified two amino acids which mediate electrostatic <i>cis</i>-interactions and analyzed the impact of disturbed <i>cis</i>-interaction on <i>trans</i>-contacts via atomic force microscopy and monitoring Förster resonance energy transfer between fluorescently labeled Cldn7-containing proteoliposomes. Our results indicate that Cldn7 <i>cis</i>-oligomerization might not be a prerequisite for establishing <i>trans</i>-contacts.
Addresses
  • Department of Chemistry, Biochemistry, Johannes Gutenberg University Mainz, Mainz, Germany.
Autoren
  • Lena Ahlswede
  • Carmen Siebenaller
  • Benedikt Junglas
  • Nadja Hellmann
  • Dirk Schneider
DOI
10.3389/fmolb.2022.908383
eISSN
2296-889X
Externe Identifier
  • PubMed Identifier: 35832741
  • PubMed Central ID: PMC9271825
Open access
true
ISSN
2296-889X
Zeitschrift
Frontiers in molecular biosciences
Sprache
eng
Medium
Electronic-eCollection
Online publication date
2022
Open access status
Open Access
Paginierung
908383
Datum der Veröffentlichung
2022
Status
Published
Publisher licence
CC BY
Datum der Datenerfassung
2022
Titel
Human Claudin-7 <i>cis</i>-Interactions Are Not Crucial for Membrane-Membrane (<i>Trans</i>-) Interactions.
Sub types
  • brief-report
  • Journal Article
Ausgabe der Zeitschrift
9

Files

https://www.frontiersin.org/articles/10.3389/fmolb.2022.908383/pdf https://europepmc.org/articles/PMC9271825?pdf=render

Data source: Europe PubMed Central

Abstract
Human Claudin-7 (Cldn7) is a member of the Claudin (Cldn) superfamily. In vivo, these proteins form tight junctions, which establish constricted connections between cells. Cldns oligomerize within the membrane plane (= cis-interaction), and also interact with Cldns from adjacent cells (= trans-interaction). Interactions of Cldns are typically studied in vivo and structural analyses of isolated Cldns are limited. Here, we describe heterologous expression in E. coli and purification of human Cldn7, enabling in vitro analyses of the isolated protein using detergent and model membrane systems. Cldn7 exists as a monomer, hexamer, and various higher oligomers in micelles. While only limited unfolding of the protein was observed in the presence of the anionic detergent sodium dodecyl sulfate, decreased ionic strength did affect Cldn7 cis-interactions. Furthermore, we identified two amino acids which mediate electrostatic cis-interactions and analyzed the impact of disturbed cis-interaction on trans-contacts via atomic force microscopy and monitoring Förster resonance energy transfer between fluorescently labeled Cldn7-containing proteoliposomes. Our results indicate that Cldn7 cis-oligomerization might not be a prerequisite for establishing trans-contacts.
Date of acceptance
2022
Autoren
  • Lena Ahlswede
  • Carmen Siebenaller
  • Benedikt Junglas
  • Nadja Hellmann
  • Dirk Schneider
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/35832741
DOI
10.3389/fmolb.2022.908383
Externe Identifier
  • PubMed Central ID: PMC9271825
ISSN
2296-889X
Zeitschrift
Front Mol Biosci
Schlüsselwörter
  • atomic force microscopy
  • claudin
  • heterologous expression
  • membrane protein
  • oligomerization
  • proteoliposomes
  • tight junction
Sprache
eng
Country
Switzerland
Paginierung
908383
PII
908383
Datum der Veröffentlichung
2022
Status
Published online
Titel
Human Claudin-7 cis-Interactions Are Not Crucial for Membrane-Membrane (Trans-) Interactions.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
9

Data source: PubMed

Author's licence
CC-BY
Autoren
  • Lena Ahlswede
  • Carmen Siebenaller
  • Benedikt Junglas
  • Nadja Hellmann
  • Dirk Schneider
Hosting institution
Universitätsbibliothek Mainz
Sammlungen
  • DFG-491381577-G
Resource version
Published version
DOI
10.3389/fmolb.2022.908383
Funding acknowledgements
  • Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 491381577
File(s) embargoed
false
Open access
true
ISSN
2296-889X
Zeitschrift
Frontiers in molecular biosciences
Schlüsselwörter
  • 570 Biowissenschaften
  • 570 Life sciences
Sprache
eng
Open access status
Open Access
Paginierung
908383
Datum der Veröffentlichung
2022
Public URL
https://openscience.ub.uni-mainz.de/handle/20.500.12030/8189
Herausgeber
Frontiers Media
Herausgeber URL
https://doi.org/10.3389/fmolb.2022.908383
Datum der Datenerfassung
2022
Datum, an dem der Datensatz öffentlich gemacht wurde
2022
Zugang
Public
Titel
Human Claudin-7 cis-interactions are not crucial for membrane-membrane (trans-) interactions
Ausgabe der Zeitschrift
9

Files

human_claudin7_cisinteraction-20221025142914201.pdf

Data source: OPENSCIENCE.UB

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