Characterization of the laminin binding domains of the Lutheran blood group glycoprotein.
Lutheran (Lu) blood group antigens and the basal cell adhesion molecule antigen reside on two glycoproteins that belong to the Ig superfamily (IgSF) and carry five Ig-like extracellular domains. These glycoproteins act as widely expressed adhesion molecules and represent the unique receptors for laminin-10/11 in erythroid cells. Here, we report the mapping of IgSF domains responsible for binding to laminin. In plasmonic resonance surface experiments, only recombinant Lu proteins containing the N-terminal IgSF domains 1-3 were able to bind laminin-10/11 and to inhibit binding of laminin to Lu-expressing K562 cells. Mutant recombinant proteins containing only IgSF domain 1, domains 1 + 2, domains 1 + 3, domains 2 + 3, domain 3, domain 4, domain 5, and domains 4 + 5 failed to bind laminin as well as a construct containing all of the extracellular domains except domain 3. Altogether, these results indicate that IgSF domains 1-3 are involved in laminin binding and that a specific spatial arrangement of these three first domains is most probably necessary for interaction. Neither the RGD nor the N-glycosylation motifs present in IgSF domain 3 were involved in laminin binding.
El Nemer
W
W
INSERM U76, Institut National de la Transfusion Sanguine, Paris 75015, France and CNRS UMR7590, Universités Paris 6 et Paris 7, Paris 75005, France.
Gane
P
P
Colin
Y
Y
D'Ambrosio
A M
AM
Callebaut
I
I
Cartron
J P
JP
Van Kim
C L
CL
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D015374:Biosensing Techniques; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D020014:K562 Cells; D007797:Laminin / Q000378:metabolism*; D008188:Lutheran Blood-Group System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D007141:Immunoglobulin Fc Fragments; D007797:Laminin; D008188:Lutheran Blood-Group System; D011993:Recombinant Fusion Proteins; C119013:laminin 10