title: Molecular determinants of S100B oligomer formation.
abstract: BACKGROUND
  S100B is a dimeric protein that can form tetramers, hexamers and higher order oligomers. These forms have been suggested to play a role in RAGE activation.
  
  
  RESULTS
  Oligomerization was found to require a low molecular weight trigger/cofactor and could not be detected for highly pure dimer, irrespective of handling. Imidazol was identified as a substance that can serve this role. Oligomerization is dependent on both the imidazol concentration and pH, with optima around 90 mM imidazol and pH 7, respectively. No oligomerization was observed above pH 8, thus the protonated form of imidazol is the active species in promoting assembly of dimers to higher species. However, disulfide bonds are not involved and the process is independent of redox potential. The process was also found to be independent of whether Ca(2+) is bound to the protein or not. Tetramers that are purified from dimers and imidazol by gel filtration are kinetically stable, but dissociate into dimers upon heating. Dimers do not revert to tetramer and higher oligomer unless imidazol is again added. Both tetramers and hexamers bind the target peptide from p53 with retained stoichiometry of one peptide per S100B monomer, and with high affinity (lgK = 7.3±0.2 and 7.2±0.2, respectively in 10 mM BisTris, 5 mM CaCl(2), pH 7.0), which is less than one order of magnitude reduced compared to dimer under the same buffer conditions.
  
  
  CONCLUSIONS
  S100B oligomerization requires protonated imidazol as a trigger/cofactor. Oligomers are kinetically stable after imidazol is removed but revert back to dimer if heated. The results underscore the importance of kinetic versus thermodynamic control of S100B protein aggregation.
authors: Department of Biochemistry, Lund University, Lund, Sweden.
authors: Eva
authors: E
authors: Thulin
authors: Tõnu
authors: T
authors: Kesvatera
authors: Sara
authors: S
authors: Linse
mesh_terms: D000595:Amino Acid Sequence; D001704:Biopolymers / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000737:chemistry*; D010084:Oxidation-Reduction; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
chemical_list: D001704:Biopolymers; D007093:Imidazoles; D009414:Nerve Growth Factors; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D012965:Sodium Chloride; D015032:Zinc; D002118:Calcium