ADP-specific sensors enable universal assay of protein kinase activity.
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.
Srinivasan
Jayaram
J
Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.
Cload
Sharon T
ST
Hamaguchi
Nobuko
N
Kurz
Jeffrey
J
Keene
Sara
S
Kurz
Markus
M
Boomer
Ryan M
RM
Blanchard
Jill
J
Epstein
David
D
Wilson
Charles
C
Diener
John L
JL
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
D008024:Ligands; D016337:RNA, Catalytic; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases