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Immobilized GSK-3 kinase and MAP kinase for modification of phosphopeptides.

Authors: Charvátová Andrea | Slováková Marcela | Korecká Lucie | Přikryl Petr | Kučerová Zdeňka | Bílková Zuzana

Year: 2009

Type of publication: ostatní - článek ve sborníku

Name of source: Amino Acids

Publisher name: Springer

Page from-to: 112-112

Titles:

Language	Name	Abstract	Keywords
cze	Imobilizovaná GSK-3 kináza a MAP kináza k modifikací fosfopeptidů	Imobilizovaná GSK-3 kináza a MAP kináza k modifikaci fosfopeptidů, pro hyperfosforylaci proteinů.	GSK-3 kinase; MAP kinase; imobilizované enzymy; magnetický nosič; fosforylace
eng	Immobilized GSK-3 kinase and MAP kinase for modification of phosphopeptides.	Attachment of phosphokinases GSK-3 kinase and MAP-kinase to magnetic microparticles carriers brings not only the advantage of an easy and repeated using but also non-contaminating and very specific and sensitive reaction on their substrates, i.e. phosphopeptides or phosphoproteins. The binding method of choice was to immobilize enzymes covalently by different methods. Reasons for failure were composed of non-oriented immobilization strategy and/or inactivation of the enzyme by reactive chemicals. As alternative proved was the attachment the enzymes tagged with His?Tag by strong affinity interaction to IMAC carrier SiMAG-IDA/Nickel. By the first method, the MALDI-TOF, we confirmed the 79 Da mass shift of the substrate in one phosphate group. For the selective detection of phosphoproteins separated by polyacrylamide gel electrophoresis the Phos-Tag Phosphoprotein Gel Stain kit has been used and method based on selective enrichment of phosphopeptides using titanium dioxide (Phos-trap).	GSK-3 kinase; MAP kinase; immobilized enzymes; magnetic carrier; phosphorylation