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Cadherin-mediated cell sorting not determined by binding or adhesion specificity.

Niessen CM, Gumbiner BM - J. Cell Biol. (2002)

Bottom Line: None of the different cadherin-expressing cells exhibited any adhesive specificity toward either of the two purified cadherin substrates, even though these cadherins differ considerably in their primary sequence.However, this lack of adhesive specificity did not determine whether different cadherin-expressing cells would sort from each other, and the tendency to sort was not predictable by the extent of sequence diversity in their extracellular domains.These results show that cadherins are far more promiscuous in their adhesive-binding capacity than had been expected and that the ability to sort out must be determined by mechanisms other than simple adhesive-binding specificity.

View Article: PubMed Central - PubMed

Affiliation: Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

ABSTRACT
Cadherin adhesion molecules play important roles in the establishment of tissue boundaries. Cells expressing different cadherins sort out from each other in cell aggregation assays. To determine the contribution of cadherin binding and adhesion specificity to the sorting process, we examined the adhesion of cells to different purified cadherin proteins. Chinese hamster ovary cell lines expressing one of four different cadherins were allowed to bind to the purified cadherin extracellular domains of either human E-cadherin or Xenopus C-cadherin, and the specificity of adhesion was compared with cell-sorting assays. None of the different cadherin-expressing cells exhibited any adhesive specificity toward either of the two purified cadherin substrates, even though these cadherins differ considerably in their primary sequence. In addition, all cells exhibited similar strengthening of adhesion on both substrates. However, this lack of adhesive specificity did not determine whether different cadherin-expressing cells would sort from each other, and the tendency to sort was not predictable by the extent of sequence diversity in their extracellular domains. These results show that cadherins are far more promiscuous in their adhesive-binding capacity than had been expected and that the ability to sort out must be determined by mechanisms other than simple adhesive-binding specificity.

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XE-cadherin CHO cells and HN-cadherin CHO cells show no adhesive specificity. (A) Adhesion flow assay of XE-CHO cells attached to either CEC1–5Fc or HEEC1–5Fc substrates (100 μg/ml). (B) Adhesion flow assay of HN-CHO attached to either CEC1–5Fc or HEEC1–5 (100 μg/ml). (C) Different concentrations of HEEC1–5Fc substrate in the adhesion flow assay using both HN-CHO and HE-CHO cells. (D) Different concentrations of CEC1–5Fc substrates in the adhesion flow assay using both HN-CHO and HE-CHO cells.
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fig5: XE-cadherin CHO cells and HN-cadherin CHO cells show no adhesive specificity. (A) Adhesion flow assay of XE-CHO cells attached to either CEC1–5Fc or HEEC1–5Fc substrates (100 μg/ml). (B) Adhesion flow assay of HN-CHO attached to either CEC1–5Fc or HEEC1–5 (100 μg/ml). (C) Different concentrations of HEEC1–5Fc substrate in the adhesion flow assay using both HN-CHO and HE-CHO cells. (D) Different concentrations of CEC1–5Fc substrates in the adhesion flow assay using both HN-CHO and HE-CHO cells.

Mentions: The adhesion of XE-CHO cells to either the human HEEC1–5Fc protein or to Xenopus CEC1–5Fc was measured. XE-CHO cells bind equally well to either CEC1–5Fc or HEEC1–5Fc in the adhesion flow assay (Fig. 5 A), again showing no specificity in adhesion to Xenopus C- or human E-cadherin. Similar to both HE-CHO cells and C-CHO cells, adhesion of XE-cadherin–expressing cells also strengthened on both cadherin substrates (unpublished data). N-cadherin is even more divergent from human E-cadherin (48% similarity; Table I) and Xenopus C-cadherin (46% similarity) and therefore might be expected to interact less well with those substrates. However, N-cadherin–expressing cells also adhered equally well to both HEEC1–5Fc and CEC1–5Fc (Fig. 5 B), and they adhered to these substrates to a similar extent as the human E-cadherin cells (Fig. 2 A). More importantly, HN-CHO and HE-CHO cells were similarly sensitive to dilution of the HEEC1–5Fc or CEC1–5Fc substrate (Fig. 5, C and D), demonstrating that differences were not being obscured by saturating levels of the substrate. In conclusion, we found that no type I classical cadherin tested thus far mediated any specificity in binding and adhesion to different purified cadherin protein substrates.


Cadherin-mediated cell sorting not determined by binding or adhesion specificity.

Niessen CM, Gumbiner BM - J. Cell Biol. (2002)

XE-cadherin CHO cells and HN-cadherin CHO cells show no adhesive specificity. (A) Adhesion flow assay of XE-CHO cells attached to either CEC1–5Fc or HEEC1–5Fc substrates (100 μg/ml). (B) Adhesion flow assay of HN-CHO attached to either CEC1–5Fc or HEEC1–5 (100 μg/ml). (C) Different concentrations of HEEC1–5Fc substrate in the adhesion flow assay using both HN-CHO and HE-CHO cells. (D) Different concentrations of CEC1–5Fc substrates in the adhesion flow assay using both HN-CHO and HE-CHO cells.
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Related In: Results  -  Collection

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fig5: XE-cadherin CHO cells and HN-cadherin CHO cells show no adhesive specificity. (A) Adhesion flow assay of XE-CHO cells attached to either CEC1–5Fc or HEEC1–5Fc substrates (100 μg/ml). (B) Adhesion flow assay of HN-CHO attached to either CEC1–5Fc or HEEC1–5 (100 μg/ml). (C) Different concentrations of HEEC1–5Fc substrate in the adhesion flow assay using both HN-CHO and HE-CHO cells. (D) Different concentrations of CEC1–5Fc substrates in the adhesion flow assay using both HN-CHO and HE-CHO cells.
Mentions: The adhesion of XE-CHO cells to either the human HEEC1–5Fc protein or to Xenopus CEC1–5Fc was measured. XE-CHO cells bind equally well to either CEC1–5Fc or HEEC1–5Fc in the adhesion flow assay (Fig. 5 A), again showing no specificity in adhesion to Xenopus C- or human E-cadherin. Similar to both HE-CHO cells and C-CHO cells, adhesion of XE-cadherin–expressing cells also strengthened on both cadherin substrates (unpublished data). N-cadherin is even more divergent from human E-cadherin (48% similarity; Table I) and Xenopus C-cadherin (46% similarity) and therefore might be expected to interact less well with those substrates. However, N-cadherin–expressing cells also adhered equally well to both HEEC1–5Fc and CEC1–5Fc (Fig. 5 B), and they adhered to these substrates to a similar extent as the human E-cadherin cells (Fig. 2 A). More importantly, HN-CHO and HE-CHO cells were similarly sensitive to dilution of the HEEC1–5Fc or CEC1–5Fc substrate (Fig. 5, C and D), demonstrating that differences were not being obscured by saturating levels of the substrate. In conclusion, we found that no type I classical cadherin tested thus far mediated any specificity in binding and adhesion to different purified cadherin protein substrates.

Bottom Line: None of the different cadherin-expressing cells exhibited any adhesive specificity toward either of the two purified cadherin substrates, even though these cadherins differ considerably in their primary sequence.However, this lack of adhesive specificity did not determine whether different cadherin-expressing cells would sort from each other, and the tendency to sort was not predictable by the extent of sequence diversity in their extracellular domains.These results show that cadherins are far more promiscuous in their adhesive-binding capacity than had been expected and that the ability to sort out must be determined by mechanisms other than simple adhesive-binding specificity.

View Article: PubMed Central - PubMed

Affiliation: Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

ABSTRACT
Cadherin adhesion molecules play important roles in the establishment of tissue boundaries. Cells expressing different cadherins sort out from each other in cell aggregation assays. To determine the contribution of cadherin binding and adhesion specificity to the sorting process, we examined the adhesion of cells to different purified cadherin proteins. Chinese hamster ovary cell lines expressing one of four different cadherins were allowed to bind to the purified cadherin extracellular domains of either human E-cadherin or Xenopus C-cadherin, and the specificity of adhesion was compared with cell-sorting assays. None of the different cadherin-expressing cells exhibited any adhesive specificity toward either of the two purified cadherin substrates, even though these cadherins differ considerably in their primary sequence. In addition, all cells exhibited similar strengthening of adhesion on both substrates. However, this lack of adhesive specificity did not determine whether different cadherin-expressing cells would sort from each other, and the tendency to sort was not predictable by the extent of sequence diversity in their extracellular domains. These results show that cadherins are far more promiscuous in their adhesive-binding capacity than had been expected and that the ability to sort out must be determined by mechanisms other than simple adhesive-binding specificity.

Show MeSH
Related in: MedlinePlus