PARP substrates and biomarkers
AssigneeBoard of Regents, The University of Texas System
InventorYonghao Yu
Life of this patent
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Abstract
PARylated proteins are enriched by treating cell lysates comprising PARylated proteins and DNA/RNA with an endonuclease that cleaves the DNA/RNA but not the PAR; and separating the PARylated proteins from the cleaved DNA/RNA. PARylation sites are labeled by eluting PARylated proteins from a PAR-affinity substrate with a nucleophilic amine exchange reactant, wherein the reactant labels PARylation sites of the proteins. Specific binding agents are identified by screening compounds for specific binding to a PARylated protein disclosed herein; and identifying one of the compounds as a specific binder of the protein. Antibodies which specifically bind PARylation sites are also disclosed.
Claims18
A method for labeling sites of poly-ADP-ribosylation (PARylation) comprising steps:
- (a) enriching for PARylated proteins comprising treating cell lysates comprising PARylated proteins and DNA/RNA with an endonuclease that cleaves the DNA/RNA but not the PAR;
- and separating the PARylated proteins from the cleaved DNA/RNA;
- (b) binding the PARylated proteins to a PAR-affinity substrate;
- and
- (c) labeling the PARylation sites comprising eluting the PARylated proteins from the PAR-affinity substrate with a nucleophilic amine exchange reactant, wherein the reactant labels PARylation sites of the proteins.
Dependencies are inferred from the claim text.