Journal article
Evaluation of a novel photoactive and biotinylated dehydroepiandrosterone analog
Molecular and cellular endocrinology, Vol.328(1), pp.56-62
2010
DOI: 10.1016/j.mce.2010.07.002
PMID: 20670672
Abstract
To characterize the cell surface receptor for dehydroepiandrosterone (DHEA), we synthesized a DHEA analog containing biotin and benzophenone groups (DHEA–BP–Bt). DHEA–BP–Bt was equipotent with DHEA in competing with [
3H]DHEA for binding to solubilized plasma membranes of bovine aortic endothelial cells (BAEC). Additionally, DHEA–BP–Bt pre-conjugated to avidin and immobilized on agarose, also inhibited plasma membrane binding of [
3H]DHEA. Furthermore, DHEA–BP–Bt activated endothelial nitric oxide synthase, similar to DHEA. Confocal micrographs showed that, upon photoirradiation, DHEA–BP–Bt bound to sites on the cell surface of BAEC in a DHEA inhibitable manner. Finally, DHEA–BP–Bt bound specifically to proteins of approximately 55
kDa and 80
kDa, either when live cells were UV irradiated with the analog and plasma membrane proteins separated by SDS-PAGE or in a ligand blot analysis. These data confirm the successful synthesis of a photoactive, biotinylated DHEA analog which is capable of cross-linking to and identifying plasma membrane DHEA binding sites and which will allow us to further purify this receptor.
Details
- Title: Subtitle
- Evaluation of a novel photoactive and biotinylated dehydroepiandrosterone analog
- Creators
- Dongmin Liu - Research Service, Department of Veterans Affairs Medical Center, Iowa City, IA, USABrianne O’Leary - Research Service, Department of Veterans Affairs Medical Center, Iowa City, IA, USAMary Iruthayanathan - Research Service, Department of Veterans Affairs Medical Center, Iowa City, IA, USALaurie Love-Homan - Division of Endocrinology, Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USANury Perez-Hernandez - Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA 52242, USAHoracio F Olivo - Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City, IA 52242, USAJoseph S Dillon - Research Service, Department of Veterans Affairs Medical Center, Iowa City, IA, USA
- Resource Type
- Journal article
- Publication Details
- Molecular and cellular endocrinology, Vol.328(1), pp.56-62
- Publisher
- Elsevier Ireland Ltd
- DOI
- 10.1016/j.mce.2010.07.002
- PMID
- 20670672
- ISSN
- 0303-7207
- eISSN
- 1872-8057
- Language
- English
- Date published
- 2010
- Academic Unit
- Surgery; Fraternal Order of Eagles Diabetes Research Center; Endocrinology and Metabolism; Internal Medicine
- Record Identifier
- 9984094772902771
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