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Galectin-4 and sulfatides in apical membrane trafficking in enterocyte-like cells.

Delacour D, Gouyer V, Zanetta JP, Drobecq H, Leteurtre E, Grard G, Moreau-Hannedouche O, Maes E, Pons A, André S, Le Bivic A, Gabius HJ, Manninen A, Simons K, Huet G - J. Cell Biol. (2005)

Bottom Line: Moreover, galectin-4 depletion altered the DRM association characteristics of apical proteins.Sulfatides with long chain-hydroxylated fatty acids, which were also enriched in DRMs, were identified as high-affinity ligands for galectin-4.Together, our data propose that interaction between galectin-4 and sulfatides plays a functional role in the clustering of lipid rafts for apical delivery.

View Article: PubMed Central - PubMed

Affiliation: Unité INSERM 560, 59045 Lille Cedex, France.

ABSTRACT
We have previously reported that 1-benzyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAc alpha-O-bn), an inhibitor of glycosylation, perturbed apical biosynthetic trafficking in polarized HT-29 cells suggesting an involvement of a lectin-based mechanism. Here, we have identified galectin-4 as one of the major components of detergent-resistant membranes (DRMs) isolated from HT-29 5M12 cells. Galectin-4 was also found in post-Golgi carrier vesicles. The functional role of galectin-4 in polarized trafficking in HT-29 5M12 cells was studied by using a retrovirus-mediated RNA interference. In galectin-4-depleted HT-29 5M12 cells apical membrane markers accumulated intracellularly. In contrast, basolateral membrane markers were not affected. Moreover, galectin-4 depletion altered the DRM association characteristics of apical proteins. Sulfatides with long chain-hydroxylated fatty acids, which were also enriched in DRMs, were identified as high-affinity ligands for galectin-4. Together, our data propose that interaction between galectin-4 and sulfatides plays a functional role in the clustering of lipid rafts for apical delivery.

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KD of galectin-4 expression in HT-29 5M12 cells induces mistargeting of apical proteins. (A) Western blot analysis of galectin-4 in empty-RVH-1-virus–infected cells or galectin-4-KD cells. (B) Confocal microscopy with antibodies directed against apical (MUC1, CEA, DPP-IV) and basolateral (E-cadherin) proteins, on empty-RVH-1-virus–infected cells or galectin-4 KD cells. xz sections are shown.
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fig6: KD of galectin-4 expression in HT-29 5M12 cells induces mistargeting of apical proteins. (A) Western blot analysis of galectin-4 in empty-RVH-1-virus–infected cells or galectin-4-KD cells. (B) Confocal microscopy with antibodies directed against apical (MUC1, CEA, DPP-IV) and basolateral (E-cadherin) proteins, on empty-RVH-1-virus–infected cells or galectin-4 KD cells. xz sections are shown.

Mentions: The level of galectin-4 mRNA depletion was analyzed by quantitative RT-PCR and Western blotting. The galectin-4 mRNA was reduced by 80% in the galectin-4-knockdown (KD) cell population and correlated well with the observed strong reduction (∼80%) of the galectin-4 protein levels (Fig. 6 A).


Galectin-4 and sulfatides in apical membrane trafficking in enterocyte-like cells.

Delacour D, Gouyer V, Zanetta JP, Drobecq H, Leteurtre E, Grard G, Moreau-Hannedouche O, Maes E, Pons A, André S, Le Bivic A, Gabius HJ, Manninen A, Simons K, Huet G - J. Cell Biol. (2005)

KD of galectin-4 expression in HT-29 5M12 cells induces mistargeting of apical proteins. (A) Western blot analysis of galectin-4 in empty-RVH-1-virus–infected cells or galectin-4-KD cells. (B) Confocal microscopy with antibodies directed against apical (MUC1, CEA, DPP-IV) and basolateral (E-cadherin) proteins, on empty-RVH-1-virus–infected cells or galectin-4 KD cells. xz sections are shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2171948&req=5

fig6: KD of galectin-4 expression in HT-29 5M12 cells induces mistargeting of apical proteins. (A) Western blot analysis of galectin-4 in empty-RVH-1-virus–infected cells or galectin-4-KD cells. (B) Confocal microscopy with antibodies directed against apical (MUC1, CEA, DPP-IV) and basolateral (E-cadherin) proteins, on empty-RVH-1-virus–infected cells or galectin-4 KD cells. xz sections are shown.
Mentions: The level of galectin-4 mRNA depletion was analyzed by quantitative RT-PCR and Western blotting. The galectin-4 mRNA was reduced by 80% in the galectin-4-knockdown (KD) cell population and correlated well with the observed strong reduction (∼80%) of the galectin-4 protein levels (Fig. 6 A).

Bottom Line: Moreover, galectin-4 depletion altered the DRM association characteristics of apical proteins.Sulfatides with long chain-hydroxylated fatty acids, which were also enriched in DRMs, were identified as high-affinity ligands for galectin-4.Together, our data propose that interaction between galectin-4 and sulfatides plays a functional role in the clustering of lipid rafts for apical delivery.

View Article: PubMed Central - PubMed

Affiliation: Unité INSERM 560, 59045 Lille Cedex, France.

ABSTRACT
We have previously reported that 1-benzyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAc alpha-O-bn), an inhibitor of glycosylation, perturbed apical biosynthetic trafficking in polarized HT-29 cells suggesting an involvement of a lectin-based mechanism. Here, we have identified galectin-4 as one of the major components of detergent-resistant membranes (DRMs) isolated from HT-29 5M12 cells. Galectin-4 was also found in post-Golgi carrier vesicles. The functional role of galectin-4 in polarized trafficking in HT-29 5M12 cells was studied by using a retrovirus-mediated RNA interference. In galectin-4-depleted HT-29 5M12 cells apical membrane markers accumulated intracellularly. In contrast, basolateral membrane markers were not affected. Moreover, galectin-4 depletion altered the DRM association characteristics of apical proteins. Sulfatides with long chain-hydroxylated fatty acids, which were also enriched in DRMs, were identified as high-affinity ligands for galectin-4. Together, our data propose that interaction between galectin-4 and sulfatides plays a functional role in the clustering of lipid rafts for apical delivery.

Show MeSH
Related in: MedlinePlus