Kinase-interacting substrate screening is a novel method to identify kinase substrates

Mutsuki Amano, Tomonari Hamaguchi, Md Hasanuzzaman Shohag, Kei Kozawa, Katsuhiro Kato, Xinjian Zhang, Yoshimitsu Yura, Yoshiharu Matsuura, Chikako Kataoka, Tomoki Nishioka, Kozo Kaibuchi

Research output: Contribution to journalArticlepeer-review

51 Citations (Scopus)


Protein kinases play pivotal roles in numerous cellular functions; however, the specific substrates of each protein kinase have not been fully elucidated. We have developed a novel method called kinase-interacting substrate screening (KISS). Using this method, 356 phosphorylation sites of 140 proteins were identified as candidate substrates for Rho-associated kinase (Rho-kinase/ROCK2), including known substrates. The KISS method was also applied to additional kinases, including PKA, MAPK1, CDK5, CaMK1, PAK7, PKN, LYN, and FYN, and a lot of candidate substrates and their phosphorylation sites were determined, most of which have not been reported previously. Among the candidate substrates for Rho-kinase, several functional clusters were identified, including the polarity-associated proteins, such as Scrib. We found that Scrib plays a crucial role in the regulation of subcellular contractility by assembling into a ternary complex with Rho-kinase and Shroom2 in a phosphorylation-dependent manner. We propose that the KISS method is a comprehensive and useful substrate screen for various kinases.

Original languageEnglish
Pages (from-to)895-912
Number of pages18
JournalJournal of Cell Biology
Issue number6
Publication statusPublished - 2015
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Cell Biology


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