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Live-Cell Assays of ER and Golgi Enzyme Activities

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R43GM105153-01A1
Agency Tracking Number: R43GM105153
Amount: $298,937.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: NIGMS
Solicitation Number: PA11-335
Timeline
Solicitation Year: 2013
Award Year: 2013
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
1300 YORK AVE, LC-523
NEW YORK, NY 10065-4805
United States
DUNS: 961919508
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 KAREN WU
 (646) 504-5697
 karen.wu@lucernatechnologies.com
Business Contact
 JENNIFER SIMONS
Phone: (646) 504-5697
Email: contact@lucernatechnologies.com
Research Institution
 Stub
Abstract

DESCRIPTION: Sirtuins and ADP ribosyltransferases (ARDTs) are two families of related enzymes that have important medically relevant functions. For example, SIRT3, a mitochondrial sirtuin, controls mitochondrial function, and thereby exerts a strong effect on metabolism, obesity, type 2 diabetes, and metabolic syndrome. There is a major need to have simple assays to measure the activity of SIRT3, as well as other sirtuins and their close relatives, the ARDTs. Each of these enzymes uses nicotinamide adeninedinucleotide (NAD), and generates nicotinamide as a byproduct of these reactions. Unfortunately, current methods for assaying the activity of these enzymes are not optimal because they are not homogeneous, are not continuous, lack sensitivity, and often not generalizable to screening all of the diverse enzymes that generate nicotinamide. A more straightforward assay would be to directly measure the products of these enzymes in a homogeneous format. Therefore, assays that directly detect the appearance of nicotinamide would be useful for measuring the activity these enzymes, regardless of the specific substrate. Using a patented sensor technology developed by Lucerna, we will develop a universal sirtuin/ARDT assay based on a simple, homogeneous fluorescencesensor that is highly specific for nicotinamide. This Phase I SBIR application describes proof-of-concept experiments for developing, characterizing, and optimizing our nicotinamide sensors in an assay for SIRT3 activity. For Phase II, our assay will be optimized to comply with standards required for ensuring that this assay can be packaged and used in a kit format for a variety of sirtuins and related enzymes. PUBLIC HEALTH RELEVANCE PUBLIC HEALTH RELEVANCE: Sirtuins and ADP ribosyltransferases are important classes of enzymes that affect many medically important processes, including cancer, inflammation, diabetes, and metabolic syndrome. Here we describe the development of a kit that can be used to assay these enzymes. This will result in an important research tool that will facilitate new discoveries about these important enzymes.

* Information listed above is at the time of submission. *

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