Kinase-interacting substrate screening is a novel method to identify kinase substrates.
Kinase-interacting substrate screening is a novel method to identify kinase substrates.
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DOI：
10.1083/jcb.201412008
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发表时间：
2015-06-22
期刊：
影响因子：
--
通讯作者：
Kaibuchi K
中科院分区：
文献类型：
--
作者：
Amano M;Hamaguchi T;Shohag MH;Kozawa K;Kato K;Zhang X;Yura Y;Matsuura Y;Kataoka C;Nishioka T;Kaibuchi K
A novel method called kinase-interacting substrate screening based on affinity beads coated with the kinase of interest identifies phosphorylation sites for Rho-kinase and others, which reveals that Rho-kinase substrate Scrib plays a crucial role in the regulation of subcellular contractility by assembling with Rho-kinase and Shroom2. 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.
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影响因子：
2.9
作者：
Amano, Mutsuki;Nakayama, Masanori;Kaibuchi, Kozo
通讯作者：
Kaibuchi, Kozo
影响因子：
14.9
作者：
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影响因子：
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作者：
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通讯作者：
Borg, JP
影响因子：
4.8
作者：
Amano, M;Ito, M;Kaibuchi, K
通讯作者：
Kaibuchi, K
影响因子：
4
作者：
Hildebrand, JD
通讯作者：
Hildebrand, JD