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PASTA: PARP activity screening and inhibitor testing assay
Ilsa T Kirby1, Daniel J Sanderson2, Michael S Cohen2
1University of California, San Francisco, Department of Cellular and Molecular Pharmacology, San Francisco, CA 94117, USA.
STAR Protocols
|March 5, 2021
Summary
Small-molecule inhibitors are crucial for studying poly-ADP-ribose polymerases (PARPs). A new assay, PARP activity screening and inhibitor testing assay (PASTA), allows researchers to determine the selectivity of PARP inhibitors against the entire enzyme family.
Area of Science:
- Biochemistry and Molecular Biology
- Enzymology
- Drug Discovery
Background:
- Poly-ADP-ribose polymerases (PARPs) are enzymes critical for various cellular processes.
- Small-molecule inhibitors have aided in understanding PARP functions.
- Limited testing of PARP inhibitors against the entire enzyme family hinders precise role assessment.
Purpose of the Study:
- To introduce a novel assay for evaluating PARP inhibitor selectivity.
- To enable confident assessment of individual PARP roles using small molecules.
Main Methods:
- Development and description of the PARP activity screening and inhibitor testing assay (PASTA).
- The assay determines the relative selectivity of various PARP inhibitors.
Main Results:
- The PASTA assay provides a method to screen PARP inhibitors across the enzyme family.
- This allows for a more accurate determination of inhibitor selectivity.
Conclusions:
- The PASTA assay is a valuable tool for researchers studying PARP enzymes and inhibitors.
- It facilitates a deeper understanding of the specific roles of individual PARPs in cellular biology.
