L-selectin, a member of the selectin family of leukocyte-endothelial adhesion proteins, mediates the initial attachment of lymphocytes to lymph node high endothelial venules during lymphocyte recirculation. One of the endothelial-associated ligands for L-selectin is GlyCAM-1, a mucin-like glycoprotein, which presents novel sulfated, sialylated and fucosylated O-glycans. In order to understand the generation of these glycans, we have examined the biosynthesis of GlyCAM-1 in lymph node organ culture. Using peptide-specific antibodies, lectins, and recombinant L-selectin, we detected the following species of GlyCAM-1: unglycosylated (<28 kDa); modified with GalNAc only (28-33 kDa); modified with sialic acid, fucose, and sulfate but lacking L-selectin reactivity (40-50 kDa); and mature (L-selectin-reactive) ligand (50-60 kDa). Pulse-chase labeling at 15 °C suggested that GalNAc is added in a pre-Golgi compartment. Treatment with brefeldin A almost completely blocked sulfation, indicating that this modification occurs in the trans-Golgi network. Two distinct sialylation events occurred in the presence of brefeldin A, while fucosylation was partially blocked. We conclude that sialylation precedes both fucosylation and sulfation during biosynthesis. This ordering will help to identify the critical acceptor structures recognized by lymph node glycosyltransferases and sulfotransferases.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				J. J Muniz, M. M Joyce, J. D Taylor II, J. R Burghardt, R. C Burghardt, and G. A Johnson Glycosylation dependent cell adhesion molecule 1-like protein and L-selectin expression in sheep interplacentomal and placentomal endometrium. Reproduction, April 1, 2006; 131(4): 751 - 761. [Abstract] [Full Text] [PDF]

				J. Mestas and C. C. W. Hughes Of Mice and Not Men: Differences between Mouse and Human Immunology J. Immunol., March 1, 2004; 172(5): 2731 - 2738. [Abstract] [Full Text] [PDF]

				X. Bai, J. R. Brown, A. Varki, and J. D. Esko Enhanced 3-O-sulfation of galactose in Asn-linked glycans and Maackia amurenesis lectin binding in a new Chinese hamster ovary cell line Glycobiology, August 1, 2001; 11(8): 621 - 632. [Abstract] [Full Text] [PDF]

				S. Hemmerich and S. D. Rosen MINI REVIEW Carbohydrate sulfotransferases in lymphocyte homing Glycobiology, September 1, 2000; 10(9): 849 - 856. [Abstract] [Full Text] [PDF]

				A. Bistrup, S. Bhakta, J. K. Lee, Y. Y. Belov, M. D. Gunn, F.-R. Zuo, C.-C. Huang, R. Kannagi, S. D. Rosen, and S. Hemmerich Sulfotransferases of Two Specificities Function in the Reconstitution of High Endothelial Cell Ligands for L-selectin J. Cell Biol., May 17, 1999; 145(4): 899 - 910. [Abstract] [Full Text] [PDF]

				T. E. Spencer, F. F. Bartol, F. W. Bazer, G. A. Johnson, and M. M. Joyce Identification and Characterization of Glycosylation-Dependent Cell Adhesion Molecule 1-Like Protein Expression in the Ovine Uterus Biol Reprod, February 1, 1999; 60(2): 241 - 250. [Abstract] [Full Text] [PDF]

				S. Degroote, J.-M. Lo-Guidice, G. Strecker, M.-P. Ducourouble, P. Roussel, and G. Lamblin Characterization of an N-Acetylglucosamine-6-O-sulfotransferase from Human Respiratory Mucosa Active on Mucin Carbohydrate Chains J. Biol. Chem., November 21, 1997; 272(47): 29493 - 29501. [Abstract] [Full Text] [PDF]

				L. Giuffre, A.-S. Cordey, N. Monai, Y. Tardy, M. Schapira, and O. Spertini Monocyte Adhesion to Activated Aortic Endothelium: Role of L-Selectin and Heparan Sulfate Proteoglycans J. Cell Biol., February 24, 1997; 136(4): 945 - 956. [Abstract] [Full Text] [PDF]

				P. L. Smith, K. M. Gersten, B. Petryniak, R. J. Kelly, C. Rogers, Y. Natsuka, J. A. Alford III, E. P. Scheidegger, S. Natsuka, and J. B. Lowe Expression of the alpha(1,3)Fucosyltransferase Fuc-TVII in Lymphoid Aggregate High Endothelial Venules Correlates with Expression of L-Selectin Ligands J. Biol. Chem., April 5, 1996; 271(14): 8250 - 8259. [Abstract] [Full Text] [PDF]

				R. E. Bruehl, C. R. Bertozzi, and S. D. Rosen Minimal Sulfated Carbohydrates for Recognition by L-selectin and the MECA-79 Antibody J. Biol. Chem., October 13, 2000; 275(42): 32642 - 32648. [Abstract] [Full Text] [PDF]