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AKT2‐signaling involves transient protein interactions
Author(s) -
Bottermann Katharina,
Reinartz Michael,
Gödecke Axel
Publication year - 2013
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.27.1_supplement.603.1
Subject(s) - akt2 , protein kinase b , chemistry , biochemistry , gene isoform , microbiology and biotechnology , signal transduction , biology , akt1 , gene
AKT is a protein kinase with widespread functions in cell metabolism, cell growth and apoptosis. Despite their high sequence homology of approx. 80 % the three AKT‐isoforms AKT 1, 2 and 3 seem to have distinct and specific functions. To analyze individual functions of the AKT2‐isoform, we examined protein interactions of AKT2 using a combination of tandem affinity purification (TAP) and quantitative mass spectrometry. AKT2 was fused with a 8 kDa TAP‐Tag and stably expressed in HEK 293 cells. Purification with AKT2‐Tag revealed Heat Shock Protein 90 (HSP 90), 78kDa glucose‐regulated protein (GRP 78), Elongation factor 2 (EF2) and Enolase 1 (ENO1) as AKT2‐interacting proteins. Using SILAC we analyzed the complex stability and discovered that AKT2 is stably bound only to the HSP 90/cdc37‐complex. Other proteins like GRP78 or Tubulin showed a low affinity binding to AKT2. In addition we could verify EF2 and ENO1 as new AKT2‐specific protein interaction partners using the proximity ligation assay (PLA) as an independent method. Taken together our data reveal that transient protein interactions play an important role for AKT2‐signaling. The newly discovered AKT2‐binding partners point to new AKT2‐mediated effects in glycolysis (ENO1) and protein synthesis (EF2).