Functional topography of myelin-associated glycoprotein. II. Mapping of domains on molecular fragments.
The myelin-associated glycoprotein (MAG), an adhesion molecule of the immunoglobulin (Ig) superfamily with five Ig-like domains, was investigated with regard to its binding site(s) for the neuronal cell surface, collagen I, and heparin, using a panel of new monoclonal antibodies and cyanogen bromide cleavage fragments of MAG. All antibodies generated competed with each other for binding to MAG, indicating that they reacted with identical or closely related epitopes. Mapping of the reactive epitopes on recombinant deletion fragments of MAG expressed by Chinese hamster ovary (CHO) fibroblasts showed reactivity of monoclonal antibody 513 with domains I, II, and III, comprising the amino-terminal end of the extracellular domain. Monoclonal antibody 15 recognized domain III only. Binding of MAG-containing liposomes to neurons was blocked by antibodies 15 and 513. Cyanogen bromide (CNBr) fragments of domains I, II, and III bound to collagen type I under isotonic buffer conditions. CNBr fragments containing domains I and II were involved in binding to heparin. These observations suggest that domain III may be important for binding to the neuronal cell surface receptor for MAG, while domains I, II, and III interact with collagen type I and domains II and III with heparin.
Meyer-Franke
A
A
Department of Neurobiology, Swiss Federal Institute of Technology, Hönggerberg, Zürich.
Tropak
M B
MB
Roder
J C
JC
Fischer
P
P
Beyreuther
K
K
Probstmeier
R
R
Schachner
M
M
D016475:3T3 Cells / Q000502:physiology; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D001665:Binding Sites / Q000502:physiology; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D016466:CHO Cells / Q000502:physiology*; D002460:Cell Line / Q000502:physiology; D003094:Collagen / Q000378:metabolism; D006224:Cricetinae; D003488:Cyanogen Bromide; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006493:Heparin / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D018992:Myelin-Associated Glycoprotein / Q000276:immunology / Q000648:ultrastructure*; D010446:Peptide Fragments
D000911:Antibodies, Monoclonal; D018992:Myelin-Associated Glycoprotein; D010446:Peptide Fragments; D006493:Heparin; D003094:Collagen; D003488:Cyanogen Bromide