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Glycogen synthase kinase 3 beta (GSK3β) forms multiple protein complexes
Author(s) -
Hamer Sabine,
Gerlach Martin,
Naguib Marian,
Reinartz Michael,
Gödecke Axel
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.465.2
Subject(s) - gsk 3 , glycogen synthase , phosphorylation , gsk3b , adenylate kinase , microbiology and biotechnology , signal transduction , biochemistry , chemistry , tandem affinity purification , wnt signaling pathway , biology , enzyme , affinity chromatography
GSK3β is a serine/threonine protein kinase, which is part of the PI3‐kinase/Akt‐, Wnt‐ and Hedgehog‐pathway and is involved in the regulation of gene transcription, protein synthesis and apoptosis. To systematically analyze GSK3β dependent signal transduction we aimed to identify GSK3β associated protein complexes via Tandem Affinity Purification (TAP) and fused a TAP‐Tag consisting of a Flag/Strep II Tag at the C‐terminal end. This enables us to perform a two‐step purification of the recombinant GSK3β and its interacting partners under native conditions. We found a similar behaviour of the recombinant GSK3β‐TAP‐Tag compared to the endogenously expressed GSK3β concerning subcellular localization, phosphorylation and enzymatic activity. MS‐based analysis of copurified proteins revealed that Acetyl‐CoA‐carboxylase, ATP‐citrate synthase and Axin, which are known to be substrates of GSK3β, form stable complexes with GSK3β. Moreover we identified numerous new interaction partners e.g. Alanyl‐tRNA synthetase, Annexin A2, Elongation factor 1, Filamin. Conclusion GSK3β mediates its signal transduction by extensive formation of stable complexes with other proteins.