Cargando…

A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase

[Image: see text] A fluorescent reagentless biosensor for ATP has been developed, based on malonyl-coenzyme A synthetase from Rhodopseudomonas palustris as the protein scaffold and recognition element. Two 5-iodoacetamidotetramethylrhodamines were covalently bound to this protein to provide the read...

Descripción completa

Detalles Bibliográficos
Autores principales: Vancraenenbroeck, Renée, Webb, Martin R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2015
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655421/
https://www.ncbi.nlm.nih.gov/pubmed/26355992
http://dx.doi.org/10.1021/acschembio.5b00346
_version_ 1782402194331402240
author Vancraenenbroeck, Renée
Webb, Martin R.
author_facet Vancraenenbroeck, Renée
Webb, Martin R.
author_sort Vancraenenbroeck, Renée
collection PubMed
description [Image: see text] A fluorescent reagentless biosensor for ATP has been developed, based on malonyl-coenzyme A synthetase from Rhodopseudomonas palustris as the protein scaffold and recognition element. Two 5-iodoacetamidotetramethylrhodamines were covalently bound to this protein to provide the readout. This adduct couples ATP binding to a 3.7-fold increase in fluorescence intensity with excitation at 553 nm and emission at 575 nm. It measures ATP concentrations with micromolar sensitivity and is highly selective for ATP relative to ADP. Its ability to monitor enzymatic ATP production or depletion was demonstrated in steady-state kinetic assays in which ATP is a product or substrate, respectively.
format Online
Article
Text
id pubmed-4655421
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-46554212015-11-27 A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase Vancraenenbroeck, Renée Webb, Martin R. ACS Chem Biol [Image: see text] A fluorescent reagentless biosensor for ATP has been developed, based on malonyl-coenzyme A synthetase from Rhodopseudomonas palustris as the protein scaffold and recognition element. Two 5-iodoacetamidotetramethylrhodamines were covalently bound to this protein to provide the readout. This adduct couples ATP binding to a 3.7-fold increase in fluorescence intensity with excitation at 553 nm and emission at 575 nm. It measures ATP concentrations with micromolar sensitivity and is highly selective for ATP relative to ADP. Its ability to monitor enzymatic ATP production or depletion was demonstrated in steady-state kinetic assays in which ATP is a product or substrate, respectively. American Chemical Society 2015-09-10 2015-11-20 /pmc/articles/PMC4655421/ /pubmed/26355992 http://dx.doi.org/10.1021/acschembio.5b00346 Text en Copyright © 2015 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Vancraenenbroeck, Renée
Webb, Martin R.
A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title_full A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title_fullStr A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title_full_unstemmed A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title_short A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
title_sort fluorescent, reagentless biosensor for atp, based on malonyl-coenzyme a synthetase
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655421/
https://www.ncbi.nlm.nih.gov/pubmed/26355992
http://dx.doi.org/10.1021/acschembio.5b00346
work_keys_str_mv AT vancraenenbroeckrenee afluorescentreagentlessbiosensorforatpbasedonmalonylcoenzymeasynthetase
AT webbmartinr afluorescentreagentlessbiosensorforatpbasedonmalonylcoenzymeasynthetase
AT vancraenenbroeckrenee fluorescentreagentlessbiosensorforatpbasedonmalonylcoenzymeasynthetase
AT webbmartinr fluorescentreagentlessbiosensorforatpbasedonmalonylcoenzymeasynthetase