Abstract: Methods of screening a test agent for activity to modulate an oxidative demethylation enzyme are provided according to embodiments of the present invention which includes preparing a test reaction comprising an oxidative demethylation enzyme, a substrate for the oxidative demethylation enzyme, a test agent and 4-amino-3-penten-2-one under conditions which allow generation of formaldehyde by oxidative demethylation of the substrate by the enzyme in the absence of the test agent; detecting fluorescence in the test reaction, the fluorescence indicative of reaction of formaldehyde and 4-amino-3-penten-2-one in the test reaction; and comparing fluorescence detected in the test reaction with a control, wherein a difference in detected fluorescence is indicative of a change in activity of the oxidative demethylation enzyme in the presence of the test agent.
Type:
Grant
Filed:
September 10, 2013
Date of Patent:
July 1, 2014
Assignee:
Arbor Assays LLC
Inventors:
Russell Hart, Barbara Scheuer, Raymond Trievel
Abstract: Methods of screening a test agent for activity to modulate an oxidative demethylation enzyme are provided according to embodiments of the present invention which includes preparing a test reaction comprising an oxidative demethylation enzyme, a substrate for the oxidative demethylation enzyme, a test agent and 4-amino-3-penten-2-one under conditions which allow generation of formaldehyde by oxidative demethylation of the substrate by the enzyme in the absence of the test agent; detecting fluorescence in the test reaction, the fluorescence indicative of reaction of formaldehyde and 4-amino-3-penten-2-one in the test reaction; and comparing fluorescence detected in the test reaction with a control, wherein a difference in detected fluorescence is indicative of a change in activity of the oxidative demethylation enzyme in the presence of the test agent.
Type:
Application
Filed:
September 10, 2013
Publication date:
January 2, 2014
Applicant:
ARBOR ASSAYS LLC
Inventors:
Russell Hart, Barbara Scheuer, Raymond Trievel
Abstract: Methods of assaying enzyme-mediated oxidative demethylation are provided according to embodiments of the present invention which includes combining, under reaction conditions, an oxidative demethylation enzyme, a substrate for the oxidative demethylation enzyme and a formaldehyde detection reagent. Detection of fluorescence is indicative of formaldehyde generated by oxidative demethylation of the substrate by the enzyme, the fluorescence resulting from reaction of formaldehyde and the formaldehyde detection reagent.
Type:
Grant
Filed:
January 14, 2009
Date of Patent:
May 8, 2012
Assignee:
Arbor Assays, LLC
Inventors:
Russell Hart, Barbara Scheuer, Raymond Trievel