Galectin-3 protein regulates mobility of N-cadherin and GM1 ganglioside at cell-cell junctions of mammary carcinoma cells.

Abstract

Galectin-3 binding to cell surface glycoproteins, including branched N-glycans generated by N-acetylglucosaminyltransferase V (Mgat5) activity, forms a multivalent, heterogeneous, and dynamic lattice. This lattice has been shown to regulate integrin and receptor tyrosine kinase signaling promoting tumor cell migration. N-cadherin is a homotypic cell-cell adhesion receptor commonly overexpressed in tumor cells that contributes to cell motility. Here we show that galectin-3 and N-cadherin interact and colocalize with the lipid raft marker GM1 ganglioside in cell-cell junctions of mammary epithelial cancer cells. Disruption of the lattice by deletion of Mgat5, siRNA depletion of galectin-3, or competitive inhibition with lactose stabilizes cell-cell junctions. It also reduces, in a p120-catenin-dependent manner, the dynamic pool of junctional N-cadherin. Proteomic analysis of detergent-resistant membranes (DRMs) revealed that the galectin lattice opposes entry of many proteins into DRM rafts. N-cadherin and catenins are present in DRMs; however, their DRM distribution is not significantly affected by lattice disruption. Galectin lattice integrity increases the mobile fraction of the raft marker, GM1 ganglioside binding cholera toxin B subunit Ctb, at cell-cell contacts in a p120-catenin-independent manner, but does not affect the mobility of either Ctb-labeled GM1 or GFP-coupled N-cadherin in nonjunctional regions. Our results suggest that the galectin lattice independently enhances lateral molecular diffusion by direct interaction with specific glycoconjugates within the adherens junction. By promoting exchange between raft and non-raft microdomains as well as molecular dynamics within junction-specific raft microdomains, the lattice may enhance turnover of N-cadherin and other glycoconjugates that determine junctional stability and rates of cell migration.

• Authors: Cécile Boscher, Yu Zi Zheng, Ramya Lakshminarayan, Ludger Johannes, James W Dennis, Leonard J Foster, Ivan R Nabi
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 287 Issue 39 Pg. 32940-52 (Sep 21 2012) ISSN: 1083-351X [Electronic] United States
• PMID: 22846995 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, CD
• CDH2 protein, human
• Cadherins
• Cdh2 protein, mouse
• Galectin 3
• Gangliosides
• Neoplasm Proteins
• ganglioside, GM4

Topics

• Animals
• Antigens, CD (genetics, metabolism)
• Breast Neoplasms (genetics, metabolism)
• Cadherins (genetics, metabolism)
• Cell Line, Tumor
• Cell Movement (genetics)
• Female
• Galectin 3 (genetics, metabolism)
• Gangliosides (genetics, metabolism)
• Humans
• Intercellular Junctions (genetics, metabolism)
• Mammary Neoplasms, Animal (genetics, metabolism)
• Membrane Microdomains (genetics, metabolism)
• Mice
• Neoplasm Proteins (genetics, metabolism)
• Protein Transport (genetics)