Preliminary crystallographic analysis of the Cks protein p13 suc1P90AP92A from Schizosacharromyces pombe

Joyanne A. Kelly, Elizabeth A. Williams, Matthew C.J. Wilce

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The p13suc1 is the fission yeast member of the Cks (Cdc28-dependant kinase subunit) family of proteins. The Cks proteins bind to and are required for the function of cyclin-dependant kinase (Cdk) proteins during cell cycle progression in eukaryotic cells. Two conformations of Cks have been detected crystallographically; a compact monomer with the C-terminal fourth β-strand inserted into the core of the molecule between strands 2 and 3, and a strand-exchanged dimer where the fourth β-strand is inserted into the core of the dimer partner in an equivalent position. There is a highly conserved "hinge" region consisting of the motif PEP, N-terminal to the fourth β-strand. In the monomer this motif constitutes a β-turn, while in the dimeric structure it is extended, allowing strand exchange. The mutant protein p13suc1P90AP92A, in which alanine residues replace both prolines of the turn, provides an opportunity to examine the role of the prolines in this hinge region and how they may allow for the formation of strand-exchanged dimers by Cks proteins. We have expressed and purified this mutant protein. Two millimolar p13suc1P90AP92A crystallised in 50 mM tris(hydroxymethyl)aminomethane pH 7.5, 30% poly(ethylene glycol) 1500. Diffraction data were collected at room temperature on an MAR345 image plate using Cu Kα radiation from a Rigaku RU200 rotating-anode generator source to 2.70Å. The crystal has unit cell parameters a=b=75.1 Å, c=34.9 Å, α=β=90°, γ=120°. Diffraction data were indexed to the space group P6 and systematic absences 00l indicate a screw axis consistent with P63.

Original languageEnglish
Pages (from-to)430-433
Number of pages4
JournalEuropean Biophysics Journal
Volume34
Issue number5
DOIs
Publication statusPublished - 1 Jul 2005
Externally publishedYes

Keywords

  • Cdk proteins
  • Cell cycle
  • Cks proteins
  • p13
  • p34

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