ABSTRACT

Basic Information

Abstract Number: 250 - 7
Author Name: Ruin Moaddel - NIA/NIH
Session Title: Chromatography Forum of the Delaware Valley Dal Nogare Award
Event Type: Awards
Event Title: The Synthesis and Characterization of SIRT6 Protein Open Tubular Column (SIRT6-OT): Characterization of the Quercetin Binding Site

Presider Name:Mary Ellen McNallyCo-Author:Nagendra Singh, Makoto Yasuda, David Wilson, Sebastian Fugmann
Affiliation:DuPont Crop ProtectionAffiliation:NIA/NIH

Date: Monday, March 18, 2013
Start Time: 10:30 AM (Slot #7)
Location: 126A

Abstract Content

SIRT6 is a histone deacetylase that has been proposed as a potential therapeutic target for metabolic disorders and the prevention of age-associated diseases. Thus the identification of compounds that modulate SIRT6 activity could be of great therapeutic importance. The aim of this study was to develop a screening method for the identification of novel modulators of SIRT6 from a natural plant extract and to characterize the binding site. To this end, we immobilized SIRT6 onto the surface of magnetic beads, and assessed SIRT6 enzymatic activity on synthetic acetylated histone tails (H3K9Ac) by measuring products of the deacetylation process. The SIRT6 coated magnetic beads were then suspended in fenugreek seed extract (Trigonella foenum-graecum) as a bait to identify active ligands suppressing SIRT6 activity. Two flavonoids, quercetin and vitexin were identified as novel modulators of SIRT6 activity.
The SIRT6 was subsequently immobilized onto the surface of an open tubular capillary to generate the SIRT6-OT column to characterize the quercetin binding site. Structurally related flavonoids were tested for activity at the SIRT6 protein, including apigenin, naringenin,luteolin and kaempferol. In addition to obtaining their binding activity using frontal affinity chromatographic techniques, we also ranked the compounds based on their ability to displace quercetin.