Profile of ligand specificity of the vitamin D binding protein for 1 alpha,25-dihydroxyvitamin D3 and its analogs.
The profile of structural preference for the ligand binding domain of the human vitamin D binding protein (DBP) was determined by steroid competition assay of 71 analogs of 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3]. The following categories of structural modification were evaluated [values represent fold change; R = reduction, I = increase in binding to the DBP from the reference 1 alpha, 25(OH)2D3]: (1) deletion in the A ring of the 1 alpha-hydroxyl-(20-1600I); (2) conversion of the triene system to the previtamin form (6-40R); (3) addition of substituents to carbon 11 of the C ring (4-14R); (4) inversion of the C/D ring junction (8-20R); (5) unsaturation of the D ring (16-ene; 4-140R); (6) replacement of hydrogen with deuterium atoms (no effect); alteration of the side chain by (7) adding or deleting carbon atoms (5-12R); (8) addition of fluorines (0.2-10R); (9) presence of unsaturation (22-ene, 0-5R; 23-ene, 3R-10I; 23-yne, 5-20R); (10) addition of hydroxyls (2-100R); and (11) addition of an aromatic ring (0-20I). Thus the DBP ligand binding domain could tolerate only modest changes to the structure of 1 alpha,25(OH)2D3 without a reduction in binding of the analog. The increases in binding seen in the aromatic side chain and with a triple bond at carbon-23 may be indicative of a preferred conformation of the flexible 1 alpha, 25(OH)2D3 side chain. In addition, a comparison was made of the DBP ligand binding domain with that of the human HL-60 cell 1 alpha, 25(OH)2D3 nuclear receptor. Both ligand binding domains could equivalently accommodate to the presence of (1) a side-chain cyclopropyl group, (2) 22-ene or 23-yne, (3) lengthening the side chain by two carbons, (4) presence of four to six fluorine atoms, (5) substitution of an oxygen for carbon 22, and (6) presence of a 22-[m-(dimethylhydroxymethyl)phenyl] aromatic group in the side chain. The DPB could tolerate better than the HL-60 cell receptor the presence of a 22-(p-hydroxyphenyl) aromatic group in the side chain and the absence of the 1 alpha-hydroxyl. In contrast, the HL-60 cell receptor could tolerate better than the DBP the following structural modifications: presence of a 16-ene, or 16-ene plus 23-yne unsaturation, and presence of an 11 beta-hydroxyl.
Department of Biochemistry, University of California, Riverside.
J E
JE
Bishop
E D
ED
Collins
W H
WH
Okamura
A W
AW
Norman
D001667:Binding, Competitive; D002117:Calcitriol / Q000031:analogs & derivatives / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D014809:Vitamin D-Binding Protein / Q000378:metabolism*
D008024:Ligands; D014809:Vitamin D-Binding Protein; D002117:Calcitriol