title: ADP-specific sensors enable universal assay of protein kinase activity.
abstract: Two molecular sensors that specifically recognize ADP in a background of over 100-fold molar excess of ATP are described. These sensors are nucleic-acid based and comprise a general method for monitoring protein kinase activity. The ADP-aptamer scintillation proximity assay is configured in a single-step, homogeneous format while the allosteric ribozyme (RiboReporter) sensor generates a fluorescent signal upon ADP-dependent ribozyme self-cleavage. Both systems perform well when configured for high-throughput screening and have been used to rediscover a known protein kinase inhibitor in a high-throughput screening format.
authors.affiliation: Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.
authors.forename: Jayaram
authors.initials: J
authors.lastname: Srinivasan
authors.forename: Sharon T
authors.initials: ST
authors.lastname: Cload
authors.forename: Nobuko
authors.initials: N
authors.lastname: Hamaguchi
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Kurz
authors.forename: Sara
authors.initials: S
authors.lastname: Keene
authors.forename: Markus
authors.initials: M
authors.lastname: Kurz
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Boomer
authors.forename: Jill
authors.initials: J
authors.lastname: Blanchard
authors.forename: David
authors.initials: D
authors.lastname: Epstein
authors.forename: Charles
authors.initials: C
authors.lastname: Wilson
authors.forename: John L
authors.initials: JL
authors.lastname: Diener
mesh_terms: D000244:Adenosine Diphosphate / Q000032:analysis* / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D008024:Ligands; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000032:analysis* / Q000378:metabolism*; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D008024:Ligands; D016337:RNA, Catalytic; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases