PUBLICATION
Chimeric Claudins: A New Tool to Study Tight Junction Structure and Function
- Authors
- Taylor, A., Warner, M., Mendoza, C., Memmott, C., LeCheminant, T., Bailey, S., Christensen, C., Keller, J., Suli, A., Mizrachi, D.
- ID
- ZDB-PUB-210622-1
- Date
- 2021
- Source
- International Journal of Molecular Sciences 22(9): (Journal)
- Registered Authors
- Keywords
- E-cadherin, claudin, membrane protein, occludin, tight junction
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Caco-2 Cells
- Cell Adhesion
- Claudins/chemistry
- Claudins/metabolism*
- Humans
- Protein Domains
- Recombinant Proteins/metabolism*
- Surface Plasmon Resonance
- Tight Junctions/chemistry*
- Tight Junctions/metabolism*
- Zebrafish
- PubMed
- 34066630 Full text @ Int. J. Mol. Sci.
Citation
Taylor, A., Warner, M., Mendoza, C., Memmott, C., LeCheminant, T., Bailey, S., Christensen, C., Keller, J., Suli, A., Mizrachi, D. (2021) Chimeric Claudins: A New Tool to Study Tight Junction Structure and Function. International Journal of Molecular Sciences. 22(9):.
Abstract
The tight junction (TJ) is a structure composed of multiple proteins, both cytosolic and membranal, responsible for cell-cell adhesion in polarized endothelium and epithelium. The TJ is intimately connected to the cytoskeleton and plays a role in development and homeostasis. Among the TJ's membrane proteins, claudins (CLDNs) are key to establishing blood-tissue barriers that protect organismal physiology. Recently, several crystal structures have been reported for detergent extracted recombinant CLDNs. These structural advances lack direct evidence to support quaternary structure of CLDNs. In this article, we have employed protein-engineering principles to create detergent-independent chimeric CLDNs, a combination of a 4-helix bundle soluble monomeric protein (PDB ID: 2jua) and the apical-50% of human CLDN1, the extracellular domain that is responsible for cell-cell adhesion. Maltose-binding protein-fused chimeric CLDNs (MBP-CCs) used in this study are soluble proteins that retain structural and functional aspects of native CLDNs. Here, we report the biophysical characterization of the structure and function of MBP-CCs. MBP-fused epithelial cadherin (MBP-eCAD) is used as a control and point of comparison of a well-characterized cell-adhesion molecule. Our synthetic strategy may benefit other families of 4-α-helix membrane proteins, including tetraspanins, connexins, pannexins, innexins, and more.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping