Journal article
Measurement of subnanomolar retinoic acid binding affinities for cellular retinoic acid binding proteins by fluorometric titration
Biochimica et biophysica acta, Protein structure and molecular enzymology, Vol.1209(1), pp.10-18
1994
DOI: 10.1016/0167-4838(94)90130-9
PMID: 7947970
Abstract
Cellular retinoic acid binding protein I (CRABP-I) and cellular retinoic acid binding protein II (CRABP-II) are small, cytoplasmic proteins which bind all-
trans-retinoic acid with high affinity. Both of these proteins belong to a family of intracellular proteins which bind amphiphilic lipids, including fatty acids, bile salts, and retinoids. Because CRABP-I and -II exhibit different tissue distributions and differential transcriptional regulation, they are proposed to serve different functions. The binding properties of mouse CRABP-I and -II purified from
Escherichia coli were examined to further understand their role in intracellular retinoic acid processing. Fluorescence titrations were performed using nanomolar protein concentrations, near the obtained dissociation constants, and analyzed by direct mathematical fitting to raw data, in order to extend the range and accuracy of binding constant determination. The apparent dissociation constants,
K′
d
, of mouse CRABP-I and CRABP-IIbinding all-
trans-retinoic acid were determined to be 0.4 ± 0.3 nM
and 2 ± 1 nM respectively, stronger binding than previously reported. The
K′
d
of mCRABP-I and mCRABP-II complexing with acitretin, a pharmacologically active synthetic retinoid used in the treatment of psoriasis, was 3 ± 1 nM and 15 ± 11nM. Both CRABPs bound 9-
cis-retinoic acid with a
K′
d of roughly 200 nM, and neither exhibited significant binding of 13-
cis-
retinoic acid
Details
- Title: Subtitle
- Measurement of subnanomolar retinoic acid binding affinities for cellular retinoic acid binding proteins by fluorometric titration
- Creators
- Andrew W. Norris - Washington University in St. LouisLin Cheng - Washington University in St. LouisVincent Giguère - Hospital for Sick ChildrenMichael Rosenberger - Hoffmann La-Roche, Inc., Nutley, NJ 07110, USAEllen Li - Washington University in St. Louis
- Resource Type
- Journal article
- Publication Details
- Biochimica et biophysica acta, Protein structure and molecular enzymology, Vol.1209(1), pp.10-18
- DOI
- 10.1016/0167-4838(94)90130-9
- PMID
- 7947970
- NLM abbreviation
- Biochim Biophys Acta
- ISSN
- 0167-4838
- eISSN
- 1879-2588
- Publisher
- Elsevier B.V
- Language
- English
- Date published
- 1994
- Academic Unit
- Endocrinology and Diabetes; Stead Family Department of Pediatrics; Biochemistry and Molecular Biology
- Record Identifier
- 9984293088802771
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