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• W4223987344 endingPage "104591" @default.
• W4223987344 startingPage "104591" @default.
• W4223987344 abstract "Phosphorylation is a posttranslational modification of proteins that regulates many cellular processes, such as communication between cells, cell proliferation, cell movements, and gene expression. Therefore, many studies have been conducted to determine the significance and function of phosphorylation. These studies involve the identification of phosphorylation site(s), kinases and phosphatases, and regulatory mechanisms. Recently, phosphorylation sites were identified using mass spectrometry and detected by immunoblotting with phosphorylation site-specific antibodies. However, the in vivo phosphorylation profile of the target protein is not easy to grasp, and the quantification of site-specific phosphorylation is challenging if the protein is phosphorylated at multiple sites. Phos-tag is a phospho-affinity SDS-PAGE approach in which phosphorylated proteins are separated depending on the number and sites of phosphorylation during electrophoresis, which overcomes the aforementioned problems. We applied this technique to perform an in vivo analysis of the phosphorylation of many proteins. In this article, we show our results for the phosphorylation of tau protein, p35 Cdk5 activator and GSK3β to reveal the utility and power of this technique in protein phosphorylation analyses in vivo. SIGNIFICANT: We show the in vivo phosphorylation of tau and two tau kinases analysed by using Phos-tag SDS-PAGE. Tau represents about 12 different phosphoisotypes when expressed in cultured cells. Tau is differently phosphorylated in patients with different tauopathy. Phosphorylation of p35 Cdk5 activator, which suppress the abnormal activation of Cdk5 by cleavage with calpain, is regulated developmentally. The Ser9 phosphorylation is not a proper marker of the GSK3β activity in vivo." @default.
• W4223987344 created "2022-04-19" @default.
• W4223987344 creator A5007629414 @default.
• W4223987344 creator A5035635053 @default.
• W4223987344 creator A5055594949 @default.
• W4223987344 date "2022-06-01" @default.
• W4223987344 modified "2023-09-26" @default.
• W4223987344 title "In vivo analysis of the phosphorylation of tau and the tau protein kinases Cdk5-p35 and GSK3β by using Phos-tag SDS–PAGE" @default.
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• W4223987344 doi "https://doi.org/10.1016/j.jprot.2022.104591" @default.
• W4223987344 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35430389" @default.
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