ADP-specific sensors enable universal assay of protein kinase activity

Jayaram Srinivasan; Sharon T Cload; Nobuko Hamaguchi; Jeffrey Kurz; Sara Keene; Markus Kurz; Ryan M Boomer; Jill Blanchard; David Epstein; Charles Wilson; John L Diener

doi:10.1016/j.chembiol.2004.03.014

ADP-specific sensors enable universal assay of protein kinase activity

Chem Biol. 2004 Apr;11(4):499-508. doi: 10.1016/j.chembiol.2004.03.014.

Authors

Jayaram Srinivasan¹, Sharon T Cload, Nobuko Hamaguchi, Jeffrey Kurz, Sara Keene, Markus Kurz, Ryan M Boomer, Jill Blanchard, David Epstein, Charles Wilson, John L Diener

Affiliation

¹ Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.

PMID: 15123244
DOI: 10.1016/j.chembiol.2004.03.014

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.

MeSH terms

Adenosine Diphosphate / analysis*
Adenosine Diphosphate / metabolism*
Adenosine Triphosphate / metabolism
Base Sequence
Biosensing Techniques / methods*
Fluorescence
Ligands
Molecular Sequence Data
Protein Kinases / analysis*
Protein Kinases / metabolism*
RNA, Catalytic / chemistry
RNA, Catalytic / metabolism
Signal Transduction
Substrate Specificity
Time Factors

Substances

Ligands
RNA, Catalytic
Adenosine Diphosphate
Adenosine Triphosphate
Protein Kinases