NMR studies of fluororetinol analogs complexed to two homologous rat cellular retinol-binding proteins.
Comparative 19F-NMR studies of fluororetinol analogs with rat cellular retinol binding protein II (CRBP II) and rat cellular retinol-binding protein (CRBP) were performed to probe differences in the binding interactions of these two homologous proteins. Line shape analyses of 19F-NMR spectra of (E,E,Z,E)-6-fluoro-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl- 2,4,6,8-nonatetren-1-ol (ligand 1), (E,E,Z,E)-6-fluoro-9-(2,2' dimethyl-6-methylcyclohexenyl)-3,7- dimethyl-2,4,6,8-nonatetren-1-ol (ligand 2), (E,Z,E,E)-5-fluoro-9-(2,2'- dimethyl-6-methylcyclohexenyl)-3,7-dimethyl-2,4,6,8-nonatetren+ ++-1-ol (ligand 3), when complexed with CRBP II at temperatures ranging from 0-45 degrees C, revealed that the 19F resonances corresponding to the bound ligand were in slow chemical exchange between two resonance frequencies. This was further supported by a 2D-NOESY exchange experiment. The kex at 25 degrees C was estimated from spectral simulation and fitting analyses to be 887 s-1, 1010 s-1 and 771 s-1 for CRBP II complexed 1, 2, and 3, respectively. In contrast, only a single absorption was observed for bound ligands complexed with rat CRBP over this temperature range, suggesting that the conformational dynamics of retinol binding are different for these two closely homologous proteins.
Rong
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Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110.
Lovey
A J
AJ
Rosenberger
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d'Avignon
D A
DA
Li
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E
D000818:Animals; D005456:Fluorescent Dyes; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D011487:Protein Conformation; D051381:Rats; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D054840:Retinol-Binding Proteins, Cellular; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014801:Vitamin A / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
D005456:Fluorescent Dyes; D008024:Ligands; C518461:Rbp2 protein, rat; D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D014801:Vitamin A