Biosynthesis of GlyCAM-1, a mucin-like ligand for L-selectin.

The Journal of Biological Chemistry
D Crommie, S D Rosen

Abstract

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 degrees 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 sulfati...Continue Reading

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Citations

Nov 10, 1998·European Journal of Immunology·V V SwarteR E Mebius
Apr 25, 1997·Journal of Immunological Methods·C R BertozziS D Rosen
Sep 16, 2000·Glycobiology·S Hemmerich, S D Rosen
Apr 6, 2006·Reproduction : the Official Journal of the Society for the Study of Fertility·Jésus J MunizGreg A Johnson
Mar 18, 2000·Leukemia & Lymphoma·S M Watt, J Y Chan
Oct 10, 2007·Analytical Biochemistry·Oliver von AhsenKarsten Parczyk
Feb 20, 1999·The Journal of Biological Chemistry·W J SandersL L Kiessling
Aug 12, 2000·The Journal of Biological Chemistry·R E BruehlS D Rosen
Mar 20, 2016·The Journal of Immunology : Official Journal of the American Association of Immunologists·David AskewAlex Y Huang
Feb 24, 2004·The Journal of Immunology : Official Journal of the American Association of Immunologists·Javier Mestas, Christopher C W Hughes
Mar 21, 1998·Biological Chemistry·A V Nieuw AmerongenE C Veerman
Feb 1, 2019·Scientific Reports·Riva Dill-GarlowAmeae M Walker

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