ArticleACS chemical biology2010
A fluorescent, reagentless biosensor for ADP based on tetramethylrhodamine-labeled ParM.
Article in ACS chemical biology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 45 citations in OpenAlex.
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- AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins.The Journal of biological chemistry · 2019Article
- Detection of Osmotic Shock-Induced Extracellular Nucleotide Release with a Genetically Encoded Fluorescent Sensor of ADP and ATP.Sensors (Basel, Switzerland) · 2019Article
- Conservation of conformational dynamics across prokaryotic actins.PLoS computational biology · 2019Article
- Conformation transitions of the polypeptide-binding pocket support an active substrate release from Hsp70s.Nature communications · 2017Article
- Development of a range of fluorescent reagentless biosensors for ATP, based on malonyl-coenzyme A synthetase.PloS one · 2017Article
- Imidazopyridazine Inhibitors of Plasmodium falciparum Calcium-Dependent Protein Kinase 1 Also Target Cyclic GMP-Dependent Protein Kinase and Heat Shock Protein 90 To Kill the Parasite at Different Stages of Intracellular Development.Antimicrobial agents and chemotherapy · 2015Article
- A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase.ACS chemical biology · 2015Article
- Biochemical and antiparasitic properties of inhibitors of the Plasmodium falciparum calcium-dependent protein kinase PfCDPK1.Antimicrobial agents and chemotherapy · 2014Article
- Protein conformational switches: from nature to design.Chemistry (Weinheim an der Bergstrasse, Germany) · 2012Review
- Detecting intramolecular conformational dynamics of single molecules in short distance range with subnanometer sensitivity.Nano letters · 2011Article
- Fluorescence detection of GDP in real time with the reagentless biosensor rhodamine-ParM.The Biochemical journal · 2011Article
- Time-resolved measurements of intracellular ATP in the yeast Saccharomyces cerevisiae using a new type of nanobiosensor.The Journal of biological chemistry · 2010Article
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2 authors.
Funding
Abstract
Fluorescence assays for ADP detection are of considerable current interest, both in basic research and in drug discovery, as they provide a generic method for measuring the activity of ATPases and kinases. The development of a novel fluorescent biosensor is described that is based on a tetramethylrhodamine-labeled, bacterial actin homologue, ParM. The design of the biosensor takes advantage of the large conformational change of ParM on ADP binding and the strong quenching of the tetramethylrhodamine fluorescence by stacking of the dye. ParM was labeled with two tetramethylrhodamines in close proximity, whereby the fluorophores are able to interact with each other. ADP binding alters the distance and relative orientation of the tetramethylrhodamines, which leads to a change in this stacking interaction and so in the fluorescence intensity. The final ADP biosensor shows approximately 15-fold fluorescence increase in response to ADP binding. It has relatively weak affinity for ADP (K(d) = 30 microM), enabling it to be used at substoichiometric concentrations relative to ADP, while reporting ADP concentration changes in a wide range around the K(d) value, namely, submicromolar to tens of micromolar. The biosensor strongly discriminates against ATP (>100-fold), allowing ADP detection against a background of millimolar ATP. At 20 degrees C, the labeled ParM binds ADP with a rate constant of 9.5 x 10(4) M(-1) s(-1) and the complex dissociates at 2.9 s(-1). Thus, the biosensor is suitable for real-time measurements, and its performance in such assays is demonstrated using a sugar kinase and a mammalian protein kinase.
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