title: Kinase-interacting substrate screening is a novel method to identify kinase substrates.
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.
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Mutsuki
authors.initials: M
authors.lastname: Amano
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Tomonari
authors.initials: T
authors.lastname: Hamaguchi
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Md Hasanuzzaman
authors.initials: MH
authors.lastname: Shohag
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Kozawa
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Kato
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Xinjian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Yura
authors.affiliation: Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Matsuura
authors.affiliation: Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Kataoka
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Nishioka
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Showa-ku, Nagoya, Aichi 466-8550, Japan kaibuchi@med.nagoya-u.ac.jp.
authors.forename: Kozo
authors.initials: K
authors.lastname: Kaibuchi
mesh_terms: D003285:Contractile Proteins / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D010766:Phosphorylation; D013379:Substrate Specificity; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D003285:Contractile Proteins; D008565:Membrane Proteins; C480591:SCRIB protein, human; D025521:Tumor Suppressor Proteins; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases