Epitope mapping of a monoclonal antibody that blocks the binding of retinol-binding protein to its receptor.
To define the receptor binding site of retinol-binding protein (RBP) we have generated monoclonal antibodies (mAbs) to human RBP and examined their ability to interfere with the receptor binding. MAbs to two conserved regions efficiently blocked the binding. No major conformational changes in the protein occurred upon mAb binding, since the mAbs could co-immunoprecipitate the RBP-transthyretin (TTR) complex. One blocking mAb showed reactivity to a synthetic peptide corresponding to one entrance loop of the retinol-binding pocket (amino acid residues 60-70). Thus, our results show that at least one of the entrance loops of the barrel of RBP is located in or close to the receptor binding site. It can also be concluded that the receptor and TTR binding sites involve different regions of RBP.
Melhus
H
H
Department of Internal Medicine, University Hospital, Uppsala, Sweden.
Båvik
C O
CO
Rask
L
L
Peterson
P A
PA
Eriksson
U
U
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001665:Binding Sites; D001667:Binding, Competitive; D018604:Epitope Mapping; D006801:Humans; D066298:In Vitro Techniques; D010455:Peptides / Q000276:immunology; D011228:Prealbumin / Q000378:metabolism*; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D010455:Peptides; D011228:Prealbumin; D011956:Receptors, Cell Surface; D012177:Retinol-Binding Proteins; C027928:retinol binding protein receptor