title: A Chimeric ATP-Linked Nucleotide Enables Luminescence Signaling of Damage Surveillance by MTH1, a Cancer Target.
abstract: The enzyme MTH1 cleanses the cellular nucleotide pool of oxidatively damaged 8-oxo-dGTP, preventing mutagenesis by this nucleotide. The enzyme is considered a promising therapeutic target; however, methods to measure its activity are indirect and laborious and have low sensitivity. Here we describe a novel ATP-linked chimeric nucleotide (ARGO) that enables luminescence signaling of the enzymatic reaction, greatly simplifying the measurement of MTH1 activity. We show that the reporting system can be used to identify inhibitors of MTH1, and we use it to quantify enzyme activity in eight cell lines and in colorectal tumor tissue. The ARGO reporter is likely to have considerable utility in the study of the biology of MTH1 and potentially in analyzing patient samples during clinical testing.
authors.affiliation: Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Debin
authors.initials: D
authors.lastname: Ji
authors.affiliation: Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Beharry
authors.affiliation: Departments of Medicine (Oncology) and Genetics, Stanford School of Medicine , Stanford, California 94305, United States.
authors.forename: James M
authors.initials: JM
authors.lastname: Ford
authors.affiliation: Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Eric T
authors.initials: ET
authors.lastname: Kool
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000378:metabolism*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D058990:Molecular Targeted Therapy; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*
chemical_list: D000255:Adenosine Triphosphate; D010744:Phosphoric Monoester Hydrolases; C078207:8-oxodGTPase; D045643:DNA Repair Enzymes