Retinoic acid is the endogenous retinoid that acts directly to support specific vitamin A-dependent processes. A long-term goal of this project is to determine whether impairment of retinoic acid biogenesis causes and/or contributes to the development of oncological and dermatological diseases that are prevented or arrested by retinoid therapy. The immediate goal is to identify and characterize the retinoid- specific oxidoreductases that catalyze the biosynthesis of retinoic acid from retinol and retinal. The hypotheses to be tested are: A) retinoic acid is generated in situ in a spectrum of tissues through the interaction of a retinol specific oxidoreductase and a low Km, low Vmax retinal dehydrogenase; B) the rate of retinoic acid synthesis is controlled by retinol availability and the activity/amount of the low Km, low Vmax retinal dehydrogenase; C) intracellular transport of retinoids occurs by direct transfer from protein to protein--e.g. retinol from cellular retinol binding protein (CRBP) to retinol dyhydrogenase, retinal from retinol dehydrogenase to retinal dehydrogenase, retinoic acid from retinal dehydrogenase to cellular retinoic acid binding protein (CRABP).
The specific aims are: 1) purify retinol and retinal dehydrogenases from rat tests cytosol; 2) characterize the pure retinoid dehydrogenases; 3) determine the tissue distribution of retinoid dehydrogenases, their concentrations in vivo and their specificity for retinoids; 4) determine whether transfer of retinoids in the metabolic pathway from retinol to retinoic acid occurs thru protein-protein complexes. The enzymes will be purified by traditional, as well as newer techniques, including affinity-, fast-protein liquid-, and immunoadsorbent chromatography. Monoclonal antibodies, raised against each dehydrogenase, will be used in enzyme-linked immunoadsorbent assays to determine tissue distribution. Kinetic experiments will be done in vitro with CRBP, CRABP and the purified dehydrogenases to determine whether protein complexes contribute to the biogenesis of retinoic acid.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK036870-04A1
Application #
3235416
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1985-07-01
Project End
1993-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost
Name
State University of New York at Buffalo
Department
Type
School of Medicine & Dentistry
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Obrochta, Kristin M; Krois, Charles R; Campos, Benito et al. (2015) Insulin regulates retinol dehydrogenase expression and all-trans-retinoic acid biosynthesis through FoxO1. J Biol Chem 290:7259-68
Obrochta, Kristin M; Kane, Maureen A; Napoli, Joseph L (2014) Effects of diet and strain on mouse serum and tissue retinoid concentrations. PLoS One 9:e99435
Chudnovskiy, Rostislav; Thompson, Airlia; Tharp, Kevin et al. (2014) Consumption of clarified grapefruit juice ameliorates high-fat diet induced insulin resistance and weight gain in mice. PLoS One 9:e108408
Ashique, Amir M; May, Scott R; Kane, Maureen A et al. (2012) Morphological defects in a novel Rdh10 mutant that has reduced retinoic acid biosynthesis and signaling. Genesis 50:415-23
Jiang, Weiya; Napoli, Joseph L (2012) Reorganization of cellular retinol-binding protein type 1 and lecithin:retinol acyltransferase during retinyl ester biosynthesis. Biochim Biophys Acta 1820:859-69
Napoli, Joseph L (2012) Physiological insights into all-trans-retinoic acid biosynthesis. Biochim Biophys Acta 1821:152-67
Wang, Chao; Kane, Maureen A; Napoli, Joseph L (2011) Multiple retinol and retinal dehydrogenases catalyze all-trans-retinoic acid biosynthesis in astrocytes. J Biol Chem 286:6542-53
Kane, Maureen A; Bright, Frank V; Napoli, Joseph L (2011) Binding affinities of CRBPI and CRBPII for 9-cis-retinoids. Biochim Biophys Acta 1810:514-8
Kane, Maureen A; Napoli, Joseph L (2010) Quantification of endogenous retinoids. Methods Mol Biol 652:1-54
Shih, Michelle Y S; Kane, Maureen A; Zhou, Ping et al. (2009) Retinol Esterification by DGAT1 Is Essential for Retinoid Homeostasis in Murine Skin. J Biol Chem 284:4292-9

Showing the most recent 10 out of 62 publications