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Improved microscale RP LC separation of phosphopeptides from in vitro phosphorylated tau protein

Authors: Kupčík Rudolf | Hromádková Lenka | Řehulka Pavel | Jankovičová Barbora | Stulík Jiří | Řípová Daniela | Bílková Zuzana

Year: 2014

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Improved microscale RP LC separation of phosphopeptides from in vitro phosphorylated tau protein	Dysfunction of tau protein, which is associated with Alzheimer's disease, can be influenced by posttranslational modifications, including increased level of site-specific phosphorylation. Hyperphosphorylation of the tau protein can result in the self-assembly of tangles of paired helical filaments and straight filaments, which are further involved in the pathogenesis of Alzheimer's disease [1]. This work was focused on in vitro phosphorylation of tau protein with soluble mitogen-activated protein kinase-1 (MAPK-1), glycogen synthase kinase 3β (GSK-3β) and cAMP-dependent protein kinase (PKA) and on detection and description of phosphorylated sites using MALDI-MS. Mixture of tau protein and used enzymes was digested with trypsin followed by TiO2 affinity purification and microscale RP LC separation. Instead of the application of highly sophisticated LC instruments, sample purification and microscale separation were done in a simple and fast protocol suitable for improved MALDI-MS detection and analysis of phosphorylated peptides.	mass spektrometry, TAU protein, phopshorylation