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Development of a HTRF® Kinase Assay for Determination of Syk Activity

Regulation of protein phosphorylation is a primary cellular signaling mechanism. Many cellular responses to internal and external events are mitigated by protein kinase signaling cascades. Dysfunction of protein kinase activity has been linked to a variety of human pathologies, in the areas of cance...

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Autores principales: Harbert, Christopher, Marshall, Jeannette, Soh, Sharon, Steger, Krista
Formato: Texto
Lenguaje:English
Publicado: Bentham Science Publishers Ltd 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774622/
https://www.ncbi.nlm.nih.gov/pubmed/20161824
http://dx.doi.org/10.2174/1875397300801010020
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author Harbert, Christopher
Marshall, Jeannette
Soh, Sharon
Steger, Krista
author_facet Harbert, Christopher
Marshall, Jeannette
Soh, Sharon
Steger, Krista
author_sort Harbert, Christopher
collection PubMed
description Regulation of protein phosphorylation is a primary cellular signaling mechanism. Many cellular responses to internal and external events are mitigated by protein kinase signaling cascades. Dysfunction of protein kinase activity has been linked to a variety of human pathologies, in the areas of cancer, inflammation, metabolism, cell cycle, apoptosis, as well as cardiovascular, neurodegenerative and autoimmune diseases [1-3]. As such, there is an important need for protein kinase activity detection methodologies for researchers engaged in Drug Discovery. A number of different technologies have been employed for the measurement of protein kinase activity, including radioactive methods, luminescent methods, and fluorescent methods. More recently, Homogeneous Time Resolved Fluorescence technology (HTRF(®)), based on the principle of time-resolved fluorescent resonance energy transfer (TR-FRET), has been developed and applied for the measurement of protein kinase activity in vitro. This technology note describes the development of an HTRF(®) assay for detection of Syk enzyme activity in a format consistent with the requirements of High-Throughput Screening (HTS) campaigns currently used in drug discovery.
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spelling pubmed-27746222010-02-16 Development of a HTRF® Kinase Assay for Determination of Syk Activity Harbert, Christopher Marshall, Jeannette Soh, Sharon Steger, Krista Curr Chem Genomics Article Regulation of protein phosphorylation is a primary cellular signaling mechanism. Many cellular responses to internal and external events are mitigated by protein kinase signaling cascades. Dysfunction of protein kinase activity has been linked to a variety of human pathologies, in the areas of cancer, inflammation, metabolism, cell cycle, apoptosis, as well as cardiovascular, neurodegenerative and autoimmune diseases [1-3]. As such, there is an important need for protein kinase activity detection methodologies for researchers engaged in Drug Discovery. A number of different technologies have been employed for the measurement of protein kinase activity, including radioactive methods, luminescent methods, and fluorescent methods. More recently, Homogeneous Time Resolved Fluorescence technology (HTRF(®)), based on the principle of time-resolved fluorescent resonance energy transfer (TR-FRET), has been developed and applied for the measurement of protein kinase activity in vitro. This technology note describes the development of an HTRF(®) assay for detection of Syk enzyme activity in a format consistent with the requirements of High-Throughput Screening (HTS) campaigns currently used in drug discovery. Bentham Science Publishers Ltd 2008-02-25 /pmc/articles/PMC2774622/ /pubmed/20161824 http://dx.doi.org/10.2174/1875397300801010020 Text en 2008 Bentham Science Publishers Ltd. http://creativecommons.org/licenses/by/2.5/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/), which permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Article
Harbert, Christopher
Marshall, Jeannette
Soh, Sharon
Steger, Krista
Development of a HTRF® Kinase Assay for Determination of Syk Activity
title Development of a HTRF® Kinase Assay for Determination of Syk Activity
title_full Development of a HTRF® Kinase Assay for Determination of Syk Activity
title_fullStr Development of a HTRF® Kinase Assay for Determination of Syk Activity
title_full_unstemmed Development of a HTRF® Kinase Assay for Determination of Syk Activity
title_short Development of a HTRF® Kinase Assay for Determination of Syk Activity
title_sort development of a htrf® kinase assay for determination of syk activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2774622/
https://www.ncbi.nlm.nih.gov/pubmed/20161824
http://dx.doi.org/10.2174/1875397300801010020
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