Reagents and Methods for Detecting Sirtuin Enzymatic Activity and Screening Sirtuin Modulators
- Detailed Technology Description
- Thisinvention provides methods and compositions for preparing a sirtuin complex anddetecting a sirtuin in a sample, as well as methods and compositions for screeningand identifying compounds which inhibit or activate the enzymatic activity of asirtuin.
- Others
-
- Mechanism-based affinity capture of sirtuins. Org. Biomol. Chemistry, 2011, 9:987-93.
- PublishedUS patent application 20130065248-A1
- *Abstract
-
Sirtuins are NAD+dependent acetyl-lysine deacetylases. Sirtuins are responsible for theregulation of a broad range of cellular processes, including energy metabolism,DNA-damage response and cellular stress resistance. The protective effects of these enzymes in age-related pathologicalprocesses, such as metabolic syndrome and neurodegenerative disorders, hasfostered a tremendous interest in studying the enzymatic activity of the sirtuins,and in the development of specific modulators of these enzymes . Thus, the development of a tool tospecifically probe sirtuin activity in biologically and pathologically relevantcontexts, as well as in screening modalities is of high practical use forassessing libraries for effects on sirtuin activity, as well as understandinghow cellular perturbations can alter sirtuin function. These are more preciseapproaches to measure sirtuin activities, rather than using downstream cellulareffects as an activity readout.
Cornell researchers have developed anovel chemical approach to rapidly isolate and detect the specific sirtuinenzymatic activities. A pan-sirtuin inhibitor reacting on sirtuin active sitesand a modified NAD+ are used to form a chemically stable complex.This complex can subsequently be cross-linked to an aldehyde-substitutedbiotin. The biotinylated sirutin complex can be retrieved by streptavidin beadsand subsequently followed by gel electrophoresis. This method enables simultaneous detection ofactive sirtuins, isolation and molecular weight determination. This method is sensitive to activity ofsirtuins.
The approach can be used to screencompounds which inhibit or activate the enzymatic activity of a sirtuin. The compounds may be useful for the studiesof sirtuins as well as for therapeutics.
PotentialApplications
- Research kits for isolate and detect the sirtuin enzymaticactivities
- Screening for compounds inhibiting or activating the enzymatic activity ofa sirtuin for research or therapeutics
Advantages
- Simultaneously and rapidly isolate and detect enzymaticactivities
- Detectenzymatic activities in a biologically and pathologically relevant context,instead of using downstream cellular effects as an activity readout
- *Licensing
- Brian J. Kellybjk44@cornell.edu212-746-6186
- Country/Region
- USA
