Cargando…

Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content

A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration...

Descripción completa

Detalles Bibliográficos
Autores principales: Kopra, Kari, Seppälä, Tanja, Rabara, Dana, Abreu-Blanco, Maria, Kulmala, Sakari, Holderfield, Matthew, Härmä, Harri
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264117/
https://www.ncbi.nlm.nih.gov/pubmed/30453509
http://dx.doi.org/10.3390/s18113989
_version_ 1783375422480711680
author Kopra, Kari
Seppälä, Tanja
Rabara, Dana
Abreu-Blanco, Maria
Kulmala, Sakari
Holderfield, Matthew
Härmä, Harri
author_facet Kopra, Kari
Seppälä, Tanja
Rabara, Dana
Abreu-Blanco, Maria
Kulmala, Sakari
Holderfield, Matthew
Härmä, Harri
author_sort Kopra, Kari
collection PubMed
description A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in their hydrolysis assays performed in a high throughput screening compatible 384-well plate format.
format Online
Article
Text
id pubmed-6264117
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-62641172018-12-12 Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content Kopra, Kari Seppälä, Tanja Rabara, Dana Abreu-Blanco, Maria Kulmala, Sakari Holderfield, Matthew Härmä, Harri Sensors (Basel) Article A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in their hydrolysis assays performed in a high throughput screening compatible 384-well plate format. MDPI 2018-11-16 /pmc/articles/PMC6264117/ /pubmed/30453509 http://dx.doi.org/10.3390/s18113989 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kopra, Kari
Seppälä, Tanja
Rabara, Dana
Abreu-Blanco, Maria
Kulmala, Sakari
Holderfield, Matthew
Härmä, Harri
Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title_full Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title_fullStr Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title_full_unstemmed Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title_short Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content
title_sort label-free time-gated luminescent detection method for the nucleotides with varying phosphate content
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264117/
https://www.ncbi.nlm.nih.gov/pubmed/30453509
http://dx.doi.org/10.3390/s18113989
work_keys_str_mv AT koprakari labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT seppalatanja labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT rabaradana labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT abreublancomaria labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT kulmalasakari labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT holderfieldmatthew labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent
AT harmaharri labelfreetimegatedluminescentdetectionmethodforthenucleotideswithvaryingphosphatecontent