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Cell surface N-glycans influence the level of functional E-cadherin at the cell-cell border.

Hall MK, Weidner DA, Dayal S, Schwalbe RA - FEBS Open Bio (2014)

Bottom Line: While it is has been established that N-glycans inside the cell impact the level of E-cadherin at the cell surface of epithelial-derived cells, it is unclear whether N-glycans outside the cell control the clustering of E-cadherin at the cell-cell border.Here, we demonstrate reduction of N-glycans at the cell surface weakened the recruitment and retention of E-cadherin at the cell-cell border, and consequently reduced the strength of cell-cell interactions.We conclude that N-glycans at the cell surface are tightly linked to the placement of E-cadherin at the cell-cell border and thereby control E-cadherin mediated cell-cell adhesion.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Brody School of Medicine at East Carolina University, Greenville, NC 27834, USA.

ABSTRACT
E-cadherin is crucial for adhesion of cells to each other and thereby development and maintenance of tissue. While it is has been established that N-glycans inside the cell impact the level of E-cadherin at the cell surface of epithelial-derived cells, it is unclear whether N-glycans outside the cell control the clustering of E-cadherin at the cell-cell border. Here, we demonstrate reduction of N-glycans at the cell surface weakened the recruitment and retention of E-cadherin at the cell-cell border, and consequently reduced the strength of cell-cell interactions. We conclude that N-glycans at the cell surface are tightly linked to the placement of E-cadherin at the cell-cell border and thereby control E-cadherin mediated cell-cell adhesion.

No MeSH data available.


Related in: MedlinePlus

Changes in levels of cell surface glycans alter E-cadherin mediated cell–cell adhesion. Typical microscopic images were acquired for E-cadherin transfected (A) and nontransfected (B) Pro-5 cells without (−, upper panel) and with (+, lower panel) PNGase F. Particles (>5 cells) of interest are encircled. Particle area (C) and particle number (D) of nontransfected and transfected CHO cells minus and plus PNGase F as indicated. Bar graphs represent data collected from 5 experiments and n ⩾ 247. One-way ANOVA with Bonferroni adjustment was used for statistical analysis. NS denotes means are not significantly different at a probability of P < 0.1.
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f0020: Changes in levels of cell surface glycans alter E-cadherin mediated cell–cell adhesion. Typical microscopic images were acquired for E-cadherin transfected (A) and nontransfected (B) Pro-5 cells without (−, upper panel) and with (+, lower panel) PNGase F. Particles (>5 cells) of interest are encircled. Particle area (C) and particle number (D) of nontransfected and transfected CHO cells minus and plus PNGase F as indicated. Bar graphs represent data collected from 5 experiments and n ⩾ 247. One-way ANOVA with Bonferroni adjustment was used for statistical analysis. NS denotes means are not significantly different at a probability of P < 0.1.

Mentions: To ascertain whether reduced levels of cell surface N-glycans correlated with lower amounts of functional E-cadherin at the cell–cell interface, we measured the strength of E-cadherin mediated cell–cell interactions. Representative images of cell dissociation assays are shown for E-cadherin transfected (Fig. 4A) and nontransfected (Fig. 4B) Pro-5 cells treated without (−) and with (+) PNGase F. Size of particles were quite similar for nontransfected Pro-5 cells undergoing PNGase F treatment while the particle area of E-cadherin transfected Pro-5 cells treated without PNGase F were about 30% larger than those cells treated with PNGase F (Fig. 4C). Further particle area of E-cadherin transfected Pro-5 cells treated with PNGase F was close to the particle area of nontransfected Pro-5 cells treated with and without PNGase F. The particle number of E-cadherin transfected Pro-5 cells treated without PNGase F was about 2 while it was about 1.5 for all other cases (Fig. 4D). These results indicate that N-glycans at the cell surface influence E-cadherin mediated cell–cell adhesion while the effect on cell–cell interactions independent of E-cadherin was negligible.


Cell surface N-glycans influence the level of functional E-cadherin at the cell-cell border.

Hall MK, Weidner DA, Dayal S, Schwalbe RA - FEBS Open Bio (2014)

Changes in levels of cell surface glycans alter E-cadherin mediated cell–cell adhesion. Typical microscopic images were acquired for E-cadherin transfected (A) and nontransfected (B) Pro-5 cells without (−, upper panel) and with (+, lower panel) PNGase F. Particles (>5 cells) of interest are encircled. Particle area (C) and particle number (D) of nontransfected and transfected CHO cells minus and plus PNGase F as indicated. Bar graphs represent data collected from 5 experiments and n ⩾ 247. One-way ANOVA with Bonferroni adjustment was used for statistical analysis. NS denotes means are not significantly different at a probability of P < 0.1.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215119&req=5

f0020: Changes in levels of cell surface glycans alter E-cadherin mediated cell–cell adhesion. Typical microscopic images were acquired for E-cadherin transfected (A) and nontransfected (B) Pro-5 cells without (−, upper panel) and with (+, lower panel) PNGase F. Particles (>5 cells) of interest are encircled. Particle area (C) and particle number (D) of nontransfected and transfected CHO cells minus and plus PNGase F as indicated. Bar graphs represent data collected from 5 experiments and n ⩾ 247. One-way ANOVA with Bonferroni adjustment was used for statistical analysis. NS denotes means are not significantly different at a probability of P < 0.1.
Mentions: To ascertain whether reduced levels of cell surface N-glycans correlated with lower amounts of functional E-cadherin at the cell–cell interface, we measured the strength of E-cadherin mediated cell–cell interactions. Representative images of cell dissociation assays are shown for E-cadherin transfected (Fig. 4A) and nontransfected (Fig. 4B) Pro-5 cells treated without (−) and with (+) PNGase F. Size of particles were quite similar for nontransfected Pro-5 cells undergoing PNGase F treatment while the particle area of E-cadherin transfected Pro-5 cells treated without PNGase F were about 30% larger than those cells treated with PNGase F (Fig. 4C). Further particle area of E-cadherin transfected Pro-5 cells treated with PNGase F was close to the particle area of nontransfected Pro-5 cells treated with and without PNGase F. The particle number of E-cadherin transfected Pro-5 cells treated without PNGase F was about 2 while it was about 1.5 for all other cases (Fig. 4D). These results indicate that N-glycans at the cell surface influence E-cadherin mediated cell–cell adhesion while the effect on cell–cell interactions independent of E-cadherin was negligible.

Bottom Line: While it is has been established that N-glycans inside the cell impact the level of E-cadherin at the cell surface of epithelial-derived cells, it is unclear whether N-glycans outside the cell control the clustering of E-cadherin at the cell-cell border.Here, we demonstrate reduction of N-glycans at the cell surface weakened the recruitment and retention of E-cadherin at the cell-cell border, and consequently reduced the strength of cell-cell interactions.We conclude that N-glycans at the cell surface are tightly linked to the placement of E-cadherin at the cell-cell border and thereby control E-cadherin mediated cell-cell adhesion.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Brody School of Medicine at East Carolina University, Greenville, NC 27834, USA.

ABSTRACT
E-cadherin is crucial for adhesion of cells to each other and thereby development and maintenance of tissue. While it is has been established that N-glycans inside the cell impact the level of E-cadherin at the cell surface of epithelial-derived cells, it is unclear whether N-glycans outside the cell control the clustering of E-cadherin at the cell-cell border. Here, we demonstrate reduction of N-glycans at the cell surface weakened the recruitment and retention of E-cadherin at the cell-cell border, and consequently reduced the strength of cell-cell interactions. We conclude that N-glycans at the cell surface are tightly linked to the placement of E-cadherin at the cell-cell border and thereby control E-cadherin mediated cell-cell adhesion.

No MeSH data available.


Related in: MedlinePlus