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ProtoArray® Protein Microarrays: Rapid Identification of PARP Substrates
Author(s) -
Nolan Edward M,
Freeman-Cook Lisa
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb542
Subject(s) - poly adp ribose polymerase , parp1 , computational biology , dna microarray , olaparib , dna damage , dna repair , biology , proteome , synthetic lethality , polymerase , proteomics , druggability , protein microarray , enzyme , biochemistry , dna , gene , gene expression
Poly(ADP‐ribose) polymerases (PARPs) play a diverse role in many physiological and pathological pathways, particularly DNA damage repair. To date, ~18 different PARP enzymes have been identified, with PARP1 being the best studied. PARP1 is activated by DNA damage and catalyzes the addition of ADP‐ribose units onto acceptor proteins, setting off the base excision repair pathway. PARP inhibitors have recently garnered attention in cancer treatment as therapeutic adjuvants used in conjunction with DNA damaging chemotherapeutics. Identifying the substrate targets of the various PARP enzymes would better define their function, help identify overlap between different PARP activities, and potentially advance clinical applications for PARP inhibitors. ProtoArray® Human Protein Microarrays contain more than 9,000 purified full‐length human proteins, providing broad coverage of the proteome for a wide range of discovery applications. We report here the development of a rapid and comprehensive assay for PARP substrate identification using ProtoArray® microarrays. Using PARP1 as a model, we identified ~110 known and novel PARP1 substrates. Our results highlight protein microarrays as a platform suitable for the global discovery of PARP substrates, enabling a more detailed understanding of their function and facilitating their potential clinical exploitation as druggable targets.