Inside-out regulation of E-cadherin conformation and adhesion

Koirala, Ramesh; Priest, Andrew Vae; Yen, Chi-Fu; Cheah, Joleen S.; Pannekoek, Willem‐Jan; Gloerich, Martijn; Yamada, Sōichiro; Sivasankar, Sanjeevi

doi:10.1073/pnas.2104090118

Cited by 32 publications

(27 citation statements)

References 78 publications

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“…47 For instance, actomyosin contractility has been shown to enhance adherens junction-based adhesion. [48][49][50] Accordingly, our Y27632 experiments might not only have reduced contractility but also adhesion. However, there would still be the disruption of the tight junctions in dKD cells.…”

Section: Discussionmentioning

confidence: 91%

Cellular segregation in co-cultures driven by differential adhesion and contractility on distinct time scales

Skamrahl¹,

Schuenemann²,

Mukenhirn

et al. 2022

Preprint

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Cellular sorting and pattern formation is crucial for many biological processes such as development, tissue regeneration, and cancer progression. Prominent physical driving forces for cellular sorting are differential adhesion and contractility. Here, we studied the segregation of epithelial co-cultures containing highly contractile, ZO1/2-depleted MDCKII cells (dKD) and their wildtype (WT) counterparts using multiple quantitative, high-throughput methods to monitor their dynamical and mechanical properties. We observe a time-dependent segregation process, governed mainly by differential contractility on short (< 5 h) and differential adhesion on long (> 5 h) time scales, respectively. The overly contractile dKD cells exert strong lateral forces on their WT neighbors, thereby apically depleting their surface area. This is reflected in a six-fold difference in excess surface area between both cell types. The lateral forces lead to a four- to six-fold increase in tension at all junctions that are in contact with the contractile cells including the interface between heterotypic cell-cell contacts. Concomitantly, the tight junction-depleted, contractile cells exhibit weaker cell-cell adhesion. Drug-induced contractility reduction delays the initial segregation but ceases to change the final demixed state, rendering differential adhesion the dominant segregation force at longer time scales. This well-controlled model system shows how cell sorting is accomplished through a complex interplay between differential adhesion and contractility and can be explained largely by generic physical driving forces.

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Section: Discussionmentioning

confidence: 91%

Cellular segregation in co-cultures driven by differential adhesion and contractility on distinct time scales

Skamrahl¹,

Schuenemann²,

Mukenhirn

et al. 2022

Preprint

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“…Our observation of X-dimers co- existing with strand-swap dimer in solution raises the possibility that a dynamic equilibrium with both states exists at cell-cell adhesion sites. Then activating antibodies could drive the dimer into the strand-swapped conformation, resulting in a strengthened state of adhesion since the strand-swap may be the more favored state under tension 43 .…”

Section: Discussionmentioning

confidence: 99%

Regulation of multiple dimeric states of E-cadherin by adhesion activating antibodies revealed through Cryo-EM and X-ray crystallography

Maker

Bolejack

Schecterson³

et al. 2022

Preprint

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E-cadherin adhesion is regulated at the cell surface, a process that can be replicated by activating antibodies. We use cryo-EM and X-ray crystallography to examine functional states of the cadherin adhesive dimer. This dimer is mediated by N-terminal beta strand-swapping involving Trp2, and forms via a different transient X-dimer intermediate. X-dimers are observed in cryo-EM along with monomers and strand-swap dimers, indicating that X-dimers form stable interactions. A novel EC4-mediated dimer was also observed. Activating Fab binding caused no gross structural changes in E-cadherin monomers but can facilitate strand swapping. Moreover, activating Fab binding is incompatible with the formation of the X-dimer. Both cryo-EM and X-ray crystallography reveal a distinctive twisted strand-swap dimer conformation caused by an outward shift in the N-terminal beta strand that may represent a strengthened state. Thus, regulation of adhesion involves changes in cadherin dimer configurations.

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“…Purified canine Ecad monomers were immobilized on AFM cantilevers (Hydra 2R-50N; AppNano) and glass coverslips (CS) as described previously 35,36 . Briefly, the CS and cantilevers were cleaned with 25% H 2 O 2 /75% H 2 SO 4 overnight and washed with Milli-Q water.…”

Section: Methodsmentioning

confidence: 99%

“…MD simulations were performed with GROMACS 2020.1 using the FARM high-performance computing cluster at University of California, Davis as described previously 35 . Briefly, the Ecad crystal structures in the absence of 19A11 (PDB ID code 2O72) and in the presence of 19A11 (PDB ID code 6CXY) were equilibrated by performing 60-ns molecular dynamics (MD) simulations.…”

Section: Methodsmentioning

confidence: 99%

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Molecular mechanism for strengthening E-cadherin adhesion using a monoclonal antibody

Xie

Maker

Priest

et al. 2022

Preprint

Self Cite

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E-cadherin (Ecad) is an essential cell-cell adhesion protein with tumor suppression properties. The adhesive state of Ecad can be modified by the monoclonal antibody 19A11, which has potential applications in reducing cancer metastasis. Using x-ray crystallography, we determine the structure of 19A11 Fab bound to Ecad and show that the antibody binds to the first extracellular domain of Ecad near its primary adhesive motif - the strand-swap dimer interface. Molecular dynamics simulations and single molecule atomic force microscopy demonstrate that 19A11 interacts with Ecad in two distinct modes, one that strengthens the strand-swap dimer and one that does not alter adhesion. We show that adhesion is strengthened by the formation of a salt bridge between 19A11 and Ecad, which in turn stabilizes the swapped β-strand and its complimentary binding pocket. Our results identify mechanistic principles for engineering antibodies to enhance Ecad adhesion.

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Inside-out regulation of E-cadherin conformation and adhesion

Cited by 32 publications

References 78 publications

Cellular segregation in co-cultures driven by differential adhesion and contractility on distinct time scales

Cellular segregation in co-cultures driven by differential adhesion and contractility on distinct time scales

Regulation of multiple dimeric states of E-cadherin by adhesion activating antibodies revealed through Cryo-EM and X-ray crystallography

Molecular mechanism for strengthening E-cadherin adhesion using a monoclonal antibody

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