title: Folding and association of the human cell cycle regulatory proteins ckshs1 and ckshs2.
abstract: The two human proteins ckshs1 and ckshs2 are each 79 amino acids in length and consist of a four-stranded beta-sheet capped at one end by two alpha-helices. They are members of the cks family of essential cell cycle regulatory proteins that can adopt two native states, a monomer and a domain-swapped dimer formed by exchange of a C-terminal beta-strand. ckshs1 and ckshs2 both have marginal thermodynamic stability (the free energies of unfolding at 25 degrees C are 3.0 and 2.5 kcal/mol, respectively) and low kinetic stability (the rates of unfolding in water are approximately 1 s(-1)). Refolding of their denatured states to the monomeric forms of the proteins is slowed by transient oligomerization that is likely to occur via domain swapping. The folding behavior of ckshs1 and ckshs2 is markedly different from that of suc1, the cks protein from Schizosaccharomyces pombe, but the domain swapping propensities are similar. The greater thermodynamic and kinetic stability of suc1 and the population of a folding intermediate are most likely a consequence of its larger size (113 residues). The similarity in the domain swapping propensities, despite the contrast in other biophysical properties, may be attributable to the common double-proline motif in the hinge loop that connects the swapped domain to the rest of the protein. The motif was shown previously for suc1 to control the equilibrium between the monomer and the domain-swapped dimer. Finally, according to our model, the kinetic barrier separating the monomer and the domain-swapped dimer arises because the protein must unfold for beta-strand exchange to occur. Consistent with this, interconversion between the two states is much faster in the human proteins than it is for suc1, reflecting the faster unfolding rates of the former.
authors.affiliation: MRC Centre for Protein Engineering, University Chemical Laboratory, Lensfield Road, Cambridge CB2 1EW, U.K.
authors.forename: Markus A
authors.initials: MA
authors.lastname: Seeliger
authors.forename: Joost W H
authors.initials: JW
authors.lastname: Schymkowitz
authors.forename: Frederic
authors.initials: F
authors.lastname: Rousseau
authors.forename: Hannah R
authors.initials: HR
authors.lastname: Wilkinson
authors.forename: Laura S
authors.initials: LS
authors.lastname: Itzhaki
mesh_terms: D042846:CDC2-CDC28 Kinases; D002152:Calorimetry, Differential Scanning; D002352:Carrier Proteins / Q000737:chemistry*; D018797:Cell Cycle Proteins; D018844:Cyclin-Dependent Kinases; D006801:Humans; D007700:Kinetics; D011487:Protein Conformation; D011494:Protein Kinases; D013816:Thermodynamics
chemical_list: C093090:CKS1B protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D011494:Protein Kinases; D042846:CDC2-CDC28 Kinases; C083689:CKS2 protein, human; D018844:Cyclin-Dependent Kinases