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AssigneeThe Board of Trustees of the University of Illinois
InventorsAmanda C. Nottbohm, Robin Shane Dothager, Karson S. Putt, Mirth T. Hoyt, Paul J. Hergenrother
Estimating the term…
Disclosed are compositions and methods capable of facilitating the detection and measurement of poly(ADP-ribose)polymerases (PARP enzymes). PARP enzyme activity can be monitored using a novel calorimetric substrate, ADP-ribose-para-nitrophenol. The substrate can be synthesized from beta nicotinamide adenine dinucleotide (β-NAD+) and para-nitrophenol. In an embodiment, a continuous assay was developed to detect and kinetically monitor activity for PARP enzymes such as PARP-1, tankyrase-1 (PARP-5), and VPARP (PARP-4). The compositions and methods are particularly useful in the screening and identification of specific PARP inhibitors.
A compound having the formula CX-1:
A method for detecting a poly(ADP-ribose) polymerase (PARP) enzyme activity, comprising providing a test sample putatively containing PARP enzymatic activity, reacting said sample with a calorimetric substrate, and observing a reaction product, thereby detecting said PARP enzyme activity.
A method of screening for a substance putatively capable of modifying a PARP enzyme activity, comprising:
A substrate compound capable of reacting specifically with a PARP enzyme, wherein said substrate is capable of forming a colorimetric product upon reaction with said PARP enzyme.
A method of synthesizing a substrate for a PARP enzyme, comprising providing a nicotinamide adenine dinucleotide component, providing a nitrophenol component, and reacting said components, thereby generating said PARP enzyme substrate.
A composition comprising compound CX-1.
A kit for detecting a presence, absence, or level of a PARP enzyme activity, comprising a PARP-specific colorimetric substrate and at least one control sample, wherein said one control sample is either a positive control sample capable of exhibiting PARP enzyme activity or a negative control sample which lacks PARP activity.
Dependencies are inferred from the claim text.