Structure-based design of small peptide inhibitors of protein kinase CK2 subunit interaction

Biochemical Journal - Tập 408 Số 3 - Trang 363-373 - 2007
Béatrice Laudet1,2,3, Caroline Barette4, Vincent Duléry5, Olivier Renaudet5, Pascal Dumy5, Alexandra Metz1,2,3, Renaud Prudent1,2,3, Alexandre Deshière1,2,3, Andréa Dessen6, Odile Filhol1,2,3, Claude Cochet1,2,3
1CEA, iRTSV/LTS, Grenoble, F-38054, France
2INSERM, U873, Grenoble F-38054, France
3Université Joseph Fourier, Grenoble, France
4CEA, iRTSV/CMBA, Grenoble, F-38054, France
5CNRS, UMR-5250, ICMG FR-2607, Grenoble, France
6CNRS, CEA, IBS/LCM, Grenoble, France

Tóm tắt

X-ray crystallography studies, as well as live-cell fluorescent imaging, have recently challenged the traditional view of protein kinase CK2. Unbalanced expression of catalytic and regulatory CK2 subunits has been observed in a variety of tissues and tumours. Thus the potential intersubunit flexibility suggested by these studies raises the likely prospect that the CK2 holoenzyme complex is subject to disassembly and reassembly. In the present paper, we show evidence for the reversible multimeric organization of the CK2 holoenzyme complex in vitro. We used a combination of site-directed mutagenesis, binding experiments and functional assays to show that, both in vitro and in vivo, only a small set of primary hydrophobic residues of CK2β which contacts at the centre of the CK2α/CK2β interface dominates affinity. The results indicate that a double mutation in CK2β of amino acids Tyr188 and Phe190, which are complementary and fill up a hydrophobic pocket of CK2α, is the most disruptive to CK2α binding both in vitro and in living cells. Further characterization of hotspots in a cluster of hydrophobic amino acids centred around Tyr188–Phe190 led us to the structure-based design of small-peptide inhibitors. One conformationally constrained 11-mer peptide (Pc) represents a unique CK2β-based small molecule that was particularly efficient (i) to antagonize the interaction between the CK2 subunits, (ii) to inhibit the assembly of the CK2 holoenzyme complex, and (iii) to strongly affect its substrate preference.

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Biochemical Journal

Tập 408 Số 3

363-373

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