Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Immobilized enzyme-based method for protein phosphorylation

Autoři: Slováková Marcela | Bílková Zuzana

Rok: 2021

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Immobilized enzyme-based method for protein phosphorylation	Protein phosphorylation has a major influence on the physiological functions of proteins, especially those that are in native conformation and involved in neurodegenerative diseases. However, structural and functional studies of these proteins require fully defined phosphorylation, including non-physiological ones. Knowledge of phosphorylation-related pathways is also essential for understanding the pathogenesis of the disease and for the designing new therapeutic strategies. Recent knowledge of kinases at molecular level, and factors affecting kinase activity and specificity, provide the opportunity to study and modulate this reaction in vitro. We can also use this knowledge for other applications. There is a huge potential to produce fully decorated recombinant proteins for biomedical or cosmetic applications, to develop safe and effective vaccines or to search for new drugs, new immunomodulators. In this work, the immobilization of immobilized serine, threonine, and tyrosine protein kinases on magnetic beads and its applications are presented. The principle of magnetic separation of particles with immobilized enzymes facilitates the laboratory comfort necessary for the separation of phosphorylated substrates. The activity and stability of the immobilized kinases were verified by using peptide substrates, and the products were analyzed by mass spectrometry, Western blotting with phosphospecific antibodies, and phosphospecific staining. Multiple phosphorylation of the model protein indicates the advantage of using immobilized kinase carriers [1]. References [1] L. Hromadkova, R. Kupcik, M. Vajrychova, P. Prikryl, A. Charvatova, B. Jankovicova, M. Slovakova, The Analyst, 143(2) (2018) 466 474.	Immobilized; enzyme-based; method; protein; phosphorylation