KANPHOS Platform: A comprehensive database for kinase-associated neural phosphorylation signaling

[Speaker] Mutsuki Amano:1
[Co-author] Junichiro Yoshimoto:2, Takayuki Kannon:3, Tomoki Nishioka:1, Shiro Usui:4, Kozo Kaibuchi:1
1:Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, Japan, 2:Graduate School of Information Science, Nara Institute of Science and Technology, Japan, 3:Faculty of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Japan, 4:Neuroinformatics Japan Center, RIKEN Brain Science Institute, Japan

Protein phosphorylation is a major and essential post-translational modification in eukaryotic cells that plays a critical role in various cellular processes. While recent advances in mass spectrometry based proteomics allowed us to identify approximately 200,000 phosphorylation sites, it is not fully understood which sites are phosphorylated by a specific kinase and which extracellular stimuli regulate the protein phosphorylation via intracellular signaling cascades. Recently, we have developed an in vitro approach termed the kinase-interacting substrate screening (KISS) method and an in vivo approach termed kinase-oriented substrate screening (KIOSS) method. Using KIOSS method, we analyzed the phosphorylation signals downstream of dopamine in mouse striatal slices, and found that about 100 proteins including ion channels and transcription factors were phosphorylated probably by PKA or MAPK. Here, we present an on-line database system which provides the phosphorylation signals identified by our KISS and KIOSS methods as well as those previously reported in the literature. The database system and its web portal, named KANPHOS (Kinase-Associated PHOspho-Signaling), were built based on the Next Generation XooNIps. All data are controlled for quality via review and curation by our professional staffs. In the portal site, we can search for the data of interest in three ways: 1) Search for substrates phosphorylated by a specific kinase; 2) Search for kinases phosphorylating a specific protein; and 3) Search for kinases and their target substrates by a specific signaling pathway. Each substrate is linked with external databases such as Uniprot KB (proteomics database), HGNC DB (human genomics database), and Allen Brain Atlas, enabling us to easily predict unknown functions of the protein phosphorylation. As an application of the database, we also demonstrate how to retrieve proteins and pathways in striatal medium-sized spiny neurons modulated by extracellular dopaminergic stimulation.

This research is supported by SRPBS and Brain/MINDS from AMED, KAKENHI from JSPS/MEXT, the Neuroinformatics Japan Center, RIKEN Brain Science institute, and the INCF Japan Node.
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