1. NMR Studies of FXR-Prenylflavonoid Complexes HHMI Summer 2009 Ryan A. Moore Dr. Victor Hsu August 25, 2009

2. Nuclear Receptors  Family of protein receptors activated by presence of steroids and hormones  Work in conjunction with other receptors to  regulate cell development and metabolism  maintain homeostasis  In presence of ligand, have ability to bind to DNA to regulate expression http://czechabsinthe.files.wordpress.com/2007/04/molecule.jpg

4. Research Focus  The farnesoid X receptor (FXR)  Expressed at high levels in liver and intestine  Bile acid-activated nuclear receptor  Contributes to bile acid synthesis and lipid metabolism  Important regulator of metabolic homeostasis http://1in100.files.wordpress.com/2009/06/dna_500.jpg

5. http:// tennesseeguy.files.wordpress.com/2008/09/hops.jpg http:// www.censcihk.com/big5/admin/web/UploadFile/200922016420253.bmp Research Focus  Xanthohumol  Found in hops (Humulus lupulus)  Prenylflavonoid  Selective bile acid receptor modulator (SBARM)  Ligands with ability to regulate distinct subset of FXR-specific genes  Widely researched for medicinal potential

6. Hypothesis Binding prenylflavonoid-type ligands to FXR results in unique structural dynamics which will yield new complexes that regulate activation of FXR target genes.

7. Project Overview  Primary approach:  Fluorescent spectroscopy to observe binding  Buffer solution containing protein is titrated with natural ligand  Compare to same solution titrated with xanthohumol

11. Continuing Research  Next step:  Label receptor with nitrogen isotope  Use nuclear magnetic resonance (NMR) spectroscopy & hydrogen/deuterium exchange mass spectroscopy http://www.pharmacy.arizona.edu/faculty/yanglab/images/NMRFacility/NMR.JPG

12. Continuing Research  From this:  Determine structure of binding site  Model how FXR binds with xanthohumol and other prenylflavonoids  Observe amide nitrogen and proton chemical shifts and determine if resulting structures are ligand specific

13. Potential Future Applications  Prenylflavonoids have not been extensively studied as specific targets for FXR receptors http://www.nassaulibrary.org/hewlett/vitamins.jpg  Pharmaceuticals containing prenylflavonoids  New treatment options for metabolic diseases  Heart disease  Type-2 diabetes  Atherosclerosis  Dietary supplements

14. Acknowledgements Dr. Victor Hsu Dr. Kevin Ahern Howard Hughes Medical Institute (HHMI)