Single-molecule analysis of cadherin-mediated cell-cell adhesion

Panorchan, Porntula; Thompson, Melissa S.; Davis, Kelly J.; Tseng, Yiider; Κωνσταντόπουλος, Κωνσταντίνος; Wirtz, Denis

doi:10.1242/jcs.02719

Cited by 196 publications

(177 citation statements)

References 56 publications

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“…6). This observation is consistent with recent findings that the kinetics of weak E-CAD12 bonds differ Ϸ30-fold from C-CAD12 bonds (11) and that human N-CAD and canine E-CAD exhibit different mechanical properties (28). However, the modest absolute differences between the bond energies should result in a distribution of homophilic and heterophilic binding, as observed in the bead assays (Fig.…”

Section: Discussionsupporting

confidence: 92%

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Similarities between heterophilic and homophilic cadherin adhesion

Prakasam

Maruthamuthu

Leckband³

2006

Proc. Natl. Acad. Sci. U.S.A.

103

107

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The mechanism that drives the segregation of cells into tissuespecific subpopulations during development is largely attributed to differences in intercellular adhesion. This process requires the cadherin family of calcium-dependent glycoproteins. A widely held view is that protein-level discrimination between different cadherins on cell surfaces drives this sorting process. Despite this postulated molecular selectivity, adhesion selectivity has not been quantitatively verified at the protein level. In this work, molecular force measurements and bead aggregation assays tested whether differences in cadherin bond strengths could account for cell sorting in vivo and in vitro. Studies were conducted with chicken N-cadherin, canine E-cadherin, and Xenopus C-cadherin. Both qualitative bead aggregation and quantitative force measurements show that the cadherins cross-react. Furthermore, heterophilic adhesion is not substantially weaker than homophilic adhesion, and the measured differences in adhesion do not correlate with cell sorting behavior. These results suggest that the basis for cell segregation during morphogenesis does not map exclusively to protein-level differences in cadherin adhesion.cell adhesion ͉ force probe ͉ selectivity ͉ surface forces apparatus B oth the formation of distinct tissues during morphogenesis and the maintenance of adult tissue structure are regulated by adhesive contacts between cells. One family of adhesion proteins, cadherins, is critical to several processes associated with the formation and maintenance of tissues. The genetic regulation of the spatiotemporal expression patterns of cadherins in vivo is believed to play a central role in embryogenesis, cell differentiation, and the maintenance of the multicellular structure of an organism (1-5). One prevalent hypothesis is that differences in adhesion between cells direct cell segregation during morphogenesis, analogous to liquid-liquid phase separation (6). A fundamental unresolved question is whether this sorting out is encoded by protein-specific or cell-specific differences in adhesion. A common hypothesis is that preferential binding between identical cadherins (homophilic) relative to heterophilic binding between dissimilar cadherins induces the cell segregation.Both the adhesive and the selectivity functions of classic cadherins map to the extracellular (EC) region (7), which comprises five tandemly arranged EC domains, EC1-5, numbered from the outermost domain (Fig. 1) (8). Classical cadherins also contain a transmembrane segment and a cytoplasmic domain. Cadherins form multiple bonds that require different EC domains (9-12). A common feature of the classical cadherins is that the N-terminal domains bind by inserting the tryptophan 2 (W2) residue from one protein into a hydrophobic pocket on an opposed N-terminal domain (13-16). However, given the weak adhesion between the outer domains (11, 12), it is questionable whether the tryptophan binding accounts for both cadherin selectivity and robust adhesion.The most extensively ci...

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Section: Discussionsupporting

confidence: 92%

“…The order of homophilic adhesion is C͞C Ͼ E͞E Ͼ N͞N, in qualitative agreement with cell adhesion and single bond rupture measurements (11,12,20,(26)(27)(28).…”

Section: Discussionsupporting

confidence: 77%

Similarities between heterophilic and homophilic cadherin adhesion

Prakasam

Maruthamuthu

Leckband³

2006

Proc. Natl. Acad. Sci. U.S.A.

103

107

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“…This value is comparable with off rates determined for other cell adhesion molecules (E-cadherin [homotypic]: k off ϭ 1.01 s Ϫ1 ; Panorchan et al, 2006; and E-selectin/carbohydrate k off ϭ 0.72 s Ϫ1 ; Tees et al, 2001) but considerably higher than the off rate determined for integrin ␣ 5 ␤ 1 -fibronectin (k off ϭ 0.012-0.13 s Ϫ1 ; Li et al, 2003). Integrin adhesion must be dynamic so that cells can respond quickly to changes in external forces.…”

Section: Discussionsupporting

confidence: 84%

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Taubenberger

Cisneros

Friedrichs

et al. 2007

MBoC

193

197

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We have characterized early steps of ␣ 2 ␤ 1 integrin-mediated cell adhesion to a collagen type I matrix by using single-cell force spectroscopy. In agreement with the role of ␣ 2 ␤ 1 as a collagen type I receptor, ␣ 2 ␤ 1 -expressing Chinese hamster ovary (CHO)-A2 cells spread rapidly on the matrix, whereas ␣ 2 ␤ 1 -negative CHO wild-type cells adhered poorly. Probing CHO-A2 cell detachment forces over a contact time range of 600 s revealed a nonlinear adhesion response. During the first 60 s, cell adhesion increased slowly, and forces associated with the smallest rupture events were consistent with the breakage of individual integrin-collagen bonds. Above 60 s, a fraction of cells rapidly switched into an activated adhesion state marked by up to 10-fold increased detachment forces. Elevated overall cell adhesion coincided with a rise of the smallest rupture forces above the value required to break a single-integrin-collagen bond, suggesting a change from single to cooperative receptor binding. Transition into the activated adhesion mode and the increase of the smallest rupture forces were both blocked by inhibitors of actomyosin contractility. We therefore propose a two-step mechanism for the establishment of ␣ 2 ␤ 1 -mediated adhesion as weak initial, single-integrin-mediated binding events are superseded by strong adhesive interactions involving receptor cooperativity and actomyosin contractility. INTRODUCTIONIntegrins are a family of ␣/␤-heterodimeric receptors involved in cell-cell adhesion and cell attachment to the extracellular matrix (Hynes, 1992). In adherent cells, integrins are often arranged into highly organized structures, such as focal complexes, focal adhesions, and fibrillar adhesions . In these mature adhesive contacts, integrins are linked to the actin cytoskeleton via their intracellular domains and a multitude of adapter proteins, such as vinculin, talin, and ␣-actinin (Zamir and Geiger, 2001). Whereas the molecular composition of integrin complexes and signaling pathways controlling their macroscopic assembly or disassembly have been analyzed in detail, less is known about the early molecular events leading to the formation of integrin-based adhesion.Generally, integrin-mediated adhesion is thought to be initiated on the cell membrane by the engagement of individual integrin dimers with their respective ligand (Lotz et al., 1989;Gallant and Garcia, 2006). The number of receptorligand pairs may then grow by increasing the cell-substrate contact area and by receptor diffusion within that zone. Subsequent adhesion strengthening occurs through integrin clustering and linkage to the cytoskeleton. In addition, the ligand affinity of the integrins may increase as a result of intracellular regulatory pathways (Hughes and Pfaff, 1998). At a later stage, the assembly of higher-order integrincytoskeletal complexes requires actomyosin-driven contractility under the control of the small guanosine triphosphatase (GTPase) RhoA (Chrzanowska-Wodnicka and Burridge, 1996) and its downstream target Rho-assoc...

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“…Epithelial cells typically express E-cadherin whereas mesenchymal cells express various other cadherins such as N-cadherin, R-cadherin, cadherin-7, or cadherin-11. Consistent with this, biophysical studies demonstrated that E-cadherin mediates stronger bonds at the molecular level than any other cadherins (Dufour et al, 1999;Chu et al, 2006;Panorchan et al, 2006). Moreover, although E-cadherin and N-cadherin may transiently coexist in certain cells, they are generally mutually exclusive, and studies on epithelial cell lines or in mouse and Xenopus embryos in which E-and N-cadherins have been interchanged clearly indicate that they can only partially substitute each other (Luo et al, 2001;Kan et al, 2007;Wheelock et al, 2008;Nandadasa et al, 2009;Libusova et al, 2010).…”

Section: Introductionmentioning

confidence: 90%

Timing and kinetics of E‐ to N‐cadherin switch during neurulation in the avian embryo

Dady

Blavet

Duband

2012

Developmental Dynamics

107

132

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Background: During embryonic development, cadherin switches are correlated with tissue remodelings, such as epithelium-to-mesenchyme transition (EMT). An E-to N-cadherin switch also occurs during neurogenesis, but this is not accompanied with EMT. The biological significance of this switch is currently unknown. Results: We analyzed the timing and kinetics of the E-to N-cadherin switch during early neural induction and neurulation in the chick embryo, in relation to the patterns of their transcriptional regulators. We found that deployment of the E-to N-cadherin switch program varies considerably along the embryonic axis. Rostrally in regions of primary neurulation, it occurs progressively both in time and space in a manner that appears neither in connection with morphological transformation of neural epithelial cells nor in synchrony with movements of neurulation. Caudally, in regions of secondary neurulation, neurogenesis was not associated with cadherin switch as N-cadherin pre-existed before formation of the neural tube. We also found that, during neural development, cadherin switch is orchestrated by a set of transcriptional regulators distinct from those involved in EMT. Conclusions: Our results indicate that cadherin switch correlates with the partition of the neurectoderm into its three main populations: ectoderm, neural crest, and neural tube.

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Single-molecule analysis of cadherin-mediated cell-cell adhesion

Cited by 196 publications

References 56 publications

Similarities between heterophilic and homophilic cadherin adhesion

Similarities between heterophilic and homophilic cadherin adhesion

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Timing and kinetics of E‐ to N‐cadherin switch during neurulation in the avian embryo

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Single-molecule analysis of cadherin-mediated cell-cell adhesion

Cited by 196 publications

References 56 publications

Similarities between heterophilic and homophilic cadherin adhesion

Similarities between heterophilic and homophilic cadherin adhesion

Revealing Early Steps of α2β1Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Timing and kinetics of E‐ to N‐cadherin switch during neurulation in the avian embryo

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Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy