Galectin-4 Binds to Sulfated Glycosphingolipids and Carcinoembryonic Antigen in Patches on the Cell Surface of Human Colon Adenocarcinoma Cells

Abstract

Galectin-4, a member of the galectin family, is expressed in the epithelium of the alimentary tract. It has two tandemly repeated carbohydrate recognition domains and specifically binds to an SO 3 ؊ 33Gal␤133GalNAc pyranoside with high affinity (Ideo, H., Seko, A., Ohkura, T., Matta, K. L., and Yamashita, K. (2002) Glycobiology 12, 199-208). In this study, we found that galectin-4 binds to glycosphingolipids carrying 3-O-sulfated Gal residues, such as SB1a, SM3, SM4s, SB2, SM2a, and GM1, but not to glycosphingolipids with 3-O-sialylated Gal, such as sLc4Cer, snLc4Cer, GM3, GM2, and GM4, using both an enzyme-linked immunosorbent assay and a surface plasmon resonance assay. A confocal immunocytochemical assay showed that galectin-4 was colocalized with SB1a, GM1, and carcinoembryonic antigen (CEA) in the patches on the cell surface of human colon adenocarcinoma CCK-81 and LS174T cells. This localization was distinct from caveolin/VIP21 localization. Furthermore, immobilized galectin-4 promoted adhesion of CCK-81 cells through the sulfated glycosphingolipid, SB1a. CEA also bound to galectin-4 with K D value of 2 ؋ 10 ؊8 M by surface plasmon resonance and coimmunoprecipitated with galectin-4 in LS174T cell lysates. These findings suggest that SB1a and CEA in the patches on the cell surface of human colon adenocarcinoma cells could be biologically important ligands for galectin-4.

P-selectin mediates metastatic progression through binding to sulfatides on tumor cells

“…The discrepancy between these results may be due to the clonal nature of the Lewis lung carcinoma cells and the subsequent differences in their signal transduction (Kabayama et al 2001;Uemura et al 2003). Recently, sulfatides were also found to be potential native ligands for Galectin-4, a galactose-binding lectin (Ideo et al 2005). Our findings propose that SM4 sulfatide is contributing to metastasis through P-selectin-mediated interactions with platelets and/or endothelium.…”

“…The lack of sulfatides resulted in neurological disorders and arrest of spermatogenesis. Several studies provided evidence that sulfatides expressed on cell surfaces of different cells exert biological functions through mediating interactions with various proteins, such as laminin, thrombospondin, amphoterin, selectins, galectin, and hepatocyte growth factor (Roberts et al 1985;Aruffo et al 1991;Suzuki et al 1993;Kobayashi et al 1994;Shikata et al 1999;Rouhiainen et al 2000;Merten and Thiagarajan 2001;Ideo et al 2005). Some of these proteins are adhesion molecules that are involved in cell -cell and cell -extracellular matrix interactions.…”

Dendritic Cell Maturation Results in Pronounced Changes in Glycan Expression Affecting Recognition by Siglecs and Galectins

“…The N-terminal CRD of galectin-8 prefers ␣2,3-sialylated galactosides as found in GM3 and GD1a, but in intact galectin-8 the two CRDs can cooperate to give highaffinity cell surface binding via LacNAc residues in N-linked glycans (31,32). The CRDs of galectin-4 bind to the 3-O-sulfated glycolipids SM4, SM3, and SB1a, but also to some glycoproteins (44). iDC express relatively high levels of the transcripts for the enzymes ␤4GalT-1, -4, and iGnT, previously described to be involved in the expression of poly-N-acetyllactosamine chains (45), which are clearly detected by MS analyses of the N-glycans.…”

Roles of galectins in inflammatory bowel disease

“…Notably, galectin-4 has been demonstrated to interact with lipid rafts of enterocytes as well, and subsequently stabilize the raft formation to generate "superrafts" [62] . A recent study has found that galectin-4 interacts with carcinoembryonic antigen of colon adenocarcinoma [63] . Alternatively, it remains obscure which glycosylated receptor(s) on intestinal CD4+ T cells is crosslinked by galectin-4.…”