A peptide microarray for detecting protein kinase activity in cell lysates.

Xiaoming Han, Yoshiki Katayama

研究成果: ジャーナルへの寄稿記事

11 引用 (Scopus)

抄録

Protein kinases (PKs) are widely recognized as valuable targets for disease diagnosis and drug discovery. For this reason, we have developed a sensitive peptide microarray for detecting intracellular PK activity. Peptides are immobilized on a glutaraldehyde-premodified high-amino terminal glass slide, by spotting 2 nL volumes of substrate peptide solutions with an automated microarray spotter. After the peptides are phosphorylated by cell lysates, phosphorylation is specifically recognized by a fluorescence-labeled antiphosphotyrosine antibody for tyrosine kinases, or Phos-tag biotin (a biotinylated phosphate-specific ligand based on Zn(2+) complex), which is subsequently bound with fluorescence-labeled streptavidin, for serine/threonine kinases. The fluorescence signal is then detected by an automatic microarray scanner. The peptide microarray system involves simple peptide immobilization, requires low sample volumes and provides a high density array. Importantly, it provides high sensitivity for detecting PK activities in cell lysates. Thus, the peptide microarray system is expected to be useful for a high-throughput kinase assay to investigate intracellular kinase activity and has potential applications in disease diagnosis and drug discovery.

元の言語英語
ページ(範囲)183-194
ページ数12
ジャーナルMethods in molecular biology (Clifton, N.J.)
669
出版物ステータス出版済み - 2010

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Protein Kinases
Peptides
Fluorescence
Drug Discovery
Phosphotransferases
Metrorrhagia
Streptavidin
Protein-Serine-Threonine Kinases
Glutaral
Biotin
Immobilization
Protein-Tyrosine Kinases
Glass
Phosphates
Phosphorylation
Ligands
Antibodies

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

これを引用

A peptide microarray for detecting protein kinase activity in cell lysates. / Han, Xiaoming; Katayama, Yoshiki.

:: Methods in molecular biology (Clifton, N.J.), 巻 669, 2010, p. 183-194.

研究成果: ジャーナルへの寄稿記事

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