PASTA: PARP activity screening and inhibitor testing assay

Ilsa T Kirby; Daniel J Sanderson; Michael S Cohen

doi:10.1016/j.xpro.2021.100344

PASTA: PARP activity screening and inhibitor testing assay

STAR Protoc. 2021 Feb 18;2(1):100344. doi: 10.1016/j.xpro.2021.100344. eCollection 2021 Mar 19.

Authors

Ilsa T Kirby¹, Daniel J Sanderson², Michael S Cohen²

Affiliations

¹ University of California, San Francisco, Department of Cellular and Molecular Pharmacology, San Francisco, CA 94117, USA.
² Oregon Health and Science University, Department of Chemical Physiology and Biochemistry, Portland, OR 97239, USA.

Abstract

Small-molecule inhibitors have been instrumental in uncovering the biological importance of poly-ADP-ribose polymerases (PARPs), a family of enzymes involved in wide-ranging aspects of cell biology. However, few PARP inhibitors are tested against the entire family of PARPs. This makes it impossible to confidently assess the role of a single PARP in cellular processes using small molecules. Here, we detail a PARP activity screening and inhibitor testing assay (PASTA) for determining relative selectivity of PARP inhibitors. For complete details on the use and execution of this protocol, please refer to Kirby et al. (2018).

Keywords: High-throughput screening; Molecular/chemical probes.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Biological Assay*
Humans
Poly(ADP-ribose) Polymerase Inhibitors / analysis*
Poly(ADP-ribose) Polymerases / chemistry*

Substances

Poly(ADP-ribose) Polymerase Inhibitors
Poly(ADP-ribose) Polymerases

Grants and funding

R01 NS088629/NS/NINDS NIH HHS/United States