Kinase-interacting substrate screening is a novel method to identify kinase substrates |
| |
| Authors: | Mutsuki Amano Tomonari Hamaguchi Md Hasanuzzaman Shohag Kei Kozawa Katsuhiro Kato Xinjian Zhang Yoshimitsu Yura Yoshiharu Matsuura Chikako Kataoka Tomoki Nishioka Kozo Kaibuchi |
| |
| Institution: | 1Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan;2Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan |
| |
| Abstract: | 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. |
| |
| Keywords: | |
|
|
