PNA-tagged Peptide Microarrays for Ratiometric Activity Detection of Cellular Protein Kinases

Hiromu IKEDA; Yoshihiro YAYAMA; Akito HATA; Jumpei KAMIMOTO; Tatsuhiro YAMAMOTO; Takeshi MORI; Yoshiki KATAYAMA

doi:10.2116/analsci.30.631

Rapid Communications

PNA-tagged Peptide Microarrays for Ratiometric Activity Detection of Cellular Protein Kinases

Hiromu IKEDA, Yoshihiro YAYAMA, Akito HATA, Jumpei KAMIMOTO, Tatsuhiro YAMAMOTO, Takeshi MORI, Yoshiki KATAYAMA

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Keywords: Protein kinase, phosphorylation, microarray, PNA, peptide, IMAC, ratiometry

JOURNAL FREE ACCESS

2014 Volume 30 Issue 6 Pages 631-635

DOI https://doi.org/10.2116/analsci.30.631

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Abstract

Peptide microarrays can be used to measure the activity of multiple protein kinases (PKs), which can be used to elucidate kinomics for drug discovery and diagnosis. Here we demonstrated a new microarray for ratiometric detection of the activity of PKs using peptide nucleic acid (PNA)-tagged peptides labeled with two different fluorophores, Cy3 and Cy5. We successfully detected cellular PK activities based on ratiometry, and applied the system for evaluation of an inhibitory drug.

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