1. Calcium-dependent dynamics of cadherin interactions at cell-cell junctions.
- Author
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Kim SA, Tai CY, Mok LP, Mosser EA, and Schuman EM
- Subjects
- Animals, COS Cells, Cadherins genetics, Calcium pharmacology, Chlorocebus aethiops, Fluorescence Resonance Energy Transfer, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, HEK293 Cells, Humans, Kinetics, L Cells, Mice, Microscopy, Confocal, Protein Binding drug effects, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Transfection, beta Catenin genetics, Cadherins metabolism, Calcium metabolism, Intercellular Junctions metabolism, beta Catenin metabolism
- Abstract
Cadherins play a key role in the dynamics of cell-cell contact formation and remodeling of junctions and tissues. Cadherin-cadherin interactions are gated by extracellular Ca(2+), which serves to rigidify the cadherin extracellular domains and promote trans junctional interactions. Here we describe the direct visualization and quantification of spatiotemporal dynamics of N-cadherin interactions across intercellular junctions in living cells using a genetically encodable FRET reporter system. Direct measurements of transjunctional cadherin interactions revealed a sudden, but partial, loss of homophilic interactions (τ = 1.17 ± 0.06 s(-1)) upon chelation of extracellular Ca(2+). A cadherin mutant with reduced adhesive activity (W2A) exhibited a faster, more substantial loss of homophilic interactions (τ = 0.86 ± 0.02 s(-1)), suggesting two types of native cadherin interactions--one that is rapidly modulated by changes in extracellular Ca(2+) and another with relatively stable adhesive activity that is Ca(2+) independent. The Ca(2+)-sensitive dynamics of cadherin interactions were transmitted to the cell interior where β-catenin translocated to N-cadherin at the junction in both cells. These data indicate that cadherins can rapidly convey dynamic information about the extracellular environment to both cells that comprise a junction.
- Published
- 2011
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