Universität zu Köln

Hubner, Gerda M., Larsen, Jane Nohr, Guerra, Barbara ORCID: 0000-0003-1136-2413, Niefind, Karsten ORCID: 0000-0002-0183-6315, Vrecl, Milka and Issinger, Olaf-Georg (2014). Evidence for aggregation of protein kinase CK2 in the cell: a novel strategy for studying CK2 holoenzyme interaction by BRET2. Mol. Cell. Biochem., 397 (1-2). S. 285 - 294. DORDRECHT: SPRINGER. ISSN 1573-4919

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Abstract

Protein kinase CK2 is a ubiquitous pro-survival kinase whose substrate targets are involved in various cellular processes. Crystal structure analysis confirmed constitutive activity of the kinase, yet CK2 activity regulation in the cell is still obscure. In-vitro studies suggest autoinhibitory aggregation of the hetero-tetrameric CK2 holoenzyme as a basis for CK2 regulation. In this study, we applied bioluminescent resonance energy transfer (BRET) technology to investigate CK2 holoenzyme aggregation in living cells. We designed a BRET2 pair consisting of the fusion proteins CK2 alpha-Rluc8 and CK2 alpha-GFP(2). This BRET2 sensor reported specific interaction of CK2 holoenzyme complexes. Furthermore, the BRET2 sensor was applied to study modulators of CK2 aggregation. We found that CK2 aggregation is not static and can be influenced by the CK2-binding protein alpha subunit of the heterotrimeric G-protein that stimulates adenylyl cyclase (G(alpha s)) and the polycationic compound polylysine. G(alpha s), but not the CK2 substrate beta-arrestin2, decreased the BRET2 signal by up to 50 %. Likewise polylysine, but not the CK2 inhibitor DRB, decreased the signal in a dose-dependent manner up to 50 %. For the first time, we present direct experimental evidence for CK2 holoenzyme aggregates in the cell. Our data suggest that CK2 activity may be controlled by holoenzyme aggregation, to our knowledge a novel mechanism for protein kinase regulation. Moreover, the BRET2 sensor used in our study is a novel tool for studying CK2 regulation by aggregation and pharmacological screening for novel allosteric CK2 effectors.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Hubner, Gerda M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Larsen, Jane Nohr UNSPECIFIED UNSPECIFIED UNSPECIFIED Guerra, Barbara UNSPECIFIED orcid.org/0000-0003-1136-2413 UNSPECIFIED Niefind, Karsten UNSPECIFIED orcid.org/0000-0002-0183-6315 UNSPECIFIED Vrecl, Milka UNSPECIFIED UNSPECIFIED UNSPECIFIED Issinger, Olaf-Georg UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-422283
DOI:	10.1007/s11010-014-2196-y
Journal or Publication Title:	Mol. Cell. Biochem.
Volume:	397
Number:	1-2
Page Range:	S. 285 - 294
Date:	2014
Publisher:	SPRINGER
Place of Publication:	DORDRECHT
ISSN:	1573-4919
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Chemistry > Institute of Biochemistry
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language REGULATORY BETA-SUBUNIT; CASEIN KINASE; BIOCHEMICAL-CHARACTERIZATION; FLUORESCENT PROTEIN; CATALYTIC SUBUNIT; CRYSTAL-STRUCTURE; SCREENING ASSAY; AUTOPHOSPHORYLATION; PHOSPHORYLATION; CK2-ALPHA Multiple languages Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/42228