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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
16
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pubmed:dateCreated |
2008-8-18
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pubmed:abstractText |
We report herein the development of an efficient fluorescence assay for serine/threonine kinases using a peptide array. Our approach is based on chemical reactions specific to phosphoserine and phosphothreonine residues, that is, base-mediated beta-elimination of the phosphate group and subsequent Michael addition of a thiol-containing fluorescent reagent. This procedure enables the covalent introduction of a fluorescent moiety into the phosphorylated peptide. Novel fluorescent reagents were designed for this purpose and synthesized. With these reagents, protein kinase A (PKA) and Akt-1 activities were readily detected. Our method can also be used to measure the activity of kinase inhibitors. This assay is expected to be widely applicable in kinase research.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
1464-3391
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:day |
15
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pubmed:volume |
16
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
7788-94
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pubmed:meshHeading |
pubmed-meshheading:18656369-Fluorescence,
pubmed-meshheading:18656369-Fluorescent Dyes,
pubmed-meshheading:18656369-Kinetics,
pubmed-meshheading:18656369-Peptides,
pubmed-meshheading:18656369-Phosphoproteins,
pubmed-meshheading:18656369-Phosphorylation,
pubmed-meshheading:18656369-Phosphoserine,
pubmed-meshheading:18656369-Phosphothreonine,
pubmed-meshheading:18656369-Protein Array Analysis,
pubmed-meshheading:18656369-Protein Kinase Inhibitors,
pubmed-meshheading:18656369-Protein-Serine-Threonine Kinases
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pubmed:year |
2008
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pubmed:articleTitle |
Array-based fluorescence assay for serine/threonine kinases using specific chemical reaction.
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pubmed:affiliation |
Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya, Aichi 467-8603, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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