Myelin-associated glycoprotein interacts with ganglioside GT1b. A mechanism for neurite outgrowth inhibition.
Myelin-associated glycoprotein (MAG) is expressed on myelinating glia and inhibits neurite outgrowth from post-natal neurons. MAG has a sialic acid binding site in its N-terminal domain and binds to specific sialylated glycans and gangliosides present on the surface of neurons, but the significance of these interactions in the effect of MAG on neurite outgrowth is unclear. Here we present evidence to suggest that recognition of sialylated glycans is essential for inhibition of neurite outgrowth by MAG. Arginine 118 on MAG is known to make a key contact with sialic acid. We show that mutation of this residue reduces the potency of MAG inhibitory activity but that residual activity is also a result of carbohydrate recognition. We then go on to investigate gangliosides GT1b and GD1a as candidate MAG receptors. We show that MAG specifically binds both gangliosides and that both are expressed on the surface of MAG-responsive neurons. Furthermore, antibody cross-linking of cell surface GT1b, but not GD1a, mimics the effect of MAG, in that neurite outgrowth is inhibited through activation of Rho kinase. These data strongly suggest that interaction with GT1b on the neuronal cell surface is a potential mechanism for inhibition of neurite outgrowth by MAG.
Neurology Centre of Excellence for Drug Discovery, GlaxoSmithKline, New Frontiers Science Park North, Third Ave., Harlow, Essex CM19 5AW, United Kingdom.
M
M
Vinson
P J
PJ
Strijbos
A
A
Rowles
L
L
Facci
S E
SE
Moore
D L
DL
Simmons
F S
FS
Walsh
D000818:Animals; D001120:Arginine / Q000378:metabolism; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D020558:GTP Phosphohydrolases / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism*; D007150:Immunohistochemistry; D018992:Myelin-Associated Glycoprotein / Q000737:chemistry / Q000378:metabolism*; D016501:Neurites; D011485:Protein Binding; D051381:Rats
D004791:Enzyme Inhibitors; D005732:Gangliosides; D018992:Myelin-Associated Glycoprotein; C016481:trisialoganglioside GT1; D001120:Arginine; D020558:GTP Phosphohydrolases