Structure-function relationships of human apolipoprotein D an immunochemical analysis.
Apolipoprotein D (apoD), a 169 amino acid member of the lipocalin family, is thought to be a transporter of small, hydrophobic ligands. A panel of 10 anti-apoD monoclonal antibodies (mAbs) was prepared and characterized in order to define apoD structure-function relationships. An apoD epitope map was constructed based on reactivity of the mAbs with apoD fragments. Three mAbs react with epitopes between apoD residues 7-78, seven mAbs with epitopes between residues 128-169, one mAb recognizes an epitope that straddles residues 99-102 and one mAb is specific for an epitope composed of non-contiguous apoD residues. Several pairs of mAbs whose respective epitopes are widely separated in apoD primary structure can compete for binding to immobilized apoD. This would be consistent with the compact beta-barrel tertiary structure that apoD is thought to adopt. None of the mAbs block the interaction of apoD with pregnenolone, a putative physiological ligand for apoD.
Terrisse
L
L
Département des Sciences Biologiques, Université du Québec à Montréal, Québec, Canada.
Marcoux
K
K
Do Carmo
S
S
Brissette
L
L
Milne
R
R
Rassart
E
E
D019138:Antibodies, Blocking / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001053:Apolipoproteins / Q000737:chemistry* / Q000276:immunology; D053399:Apolipoproteins D; D001667:Binding, Competitive; D015415:Biomarkers / Q000737:chemistry*; D018604:Epitope Mapping; D006801:Humans; D007120:Immunochemistry / Q000379:methods*; D008024:Ligands; D011284:Pregnenolone / Q000276:immunology; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000276:immunology
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D001053:Apolipoproteins; D053399:Apolipoproteins D; D015415:Biomarkers; D008024:Ligands; D011993:Recombinant Fusion Proteins; D011284:Pregnenolone; D001616:beta-Galactosidase