title: An electrospray-ionization mass spectrometry analysis of the pH-dependent dissociation and denaturation processes of a heterodimeric protein.
abstract: Electrospray ionization mass spectrometry (ESI-MS) was applied to the analysis of the dissociation and denaturation processes of a heterodimeric yeast killer toxin SMKT. The two distinct subunits of SMKT noncovalently associate under acidic conditions, but become dissociated and denatured under neutral and basic conditions. In order to understand the unique pH-dependent denaturation mechanism of this protein, a pH titration was performed by utilizing ESI-MS. The molecular ions of the heterodimer which possesses the highly ordered structure, were mainly observed below pH 4.6. However, the two subunits immediately dissociated at this pH. The spectra measured with various settings of the mass spectrometer indirectly demonstrated that the pH-dependent dissociation occurs in the liquid phase. The current result as well as the three-dimensional structure of SMKT suggest that the deprotonation of a specific carboxyl group triggers a cooperative dissociation process of this protein. In conclusion, the pH titration of a protein by ESI-MS is particularly effective, when the unfolding process or the biological function of the protein is related to the interaction with other molecules.
authors.affiliation: Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Kanagawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kashiwagi
authors.forename: N
authors.initials: N
authors.lastname: Yamada
authors.forename: K
authors.initials: K
authors.lastname: Hirayama
authors.forename: C
authors.initials: C
authors.lastname: Suzuki
authors.forename: Y
authors.initials: Y
authors.lastname: Kashiwagi
authors.forename: F
authors.initials: F
authors.lastname: Tsuchiya
authors.forename: Y
authors.initials: Y
authors.lastname: Arata
authors.forename: N
authors.initials: N
authors.lastname: Kunishima
authors.forename: K
authors.initials: K
authors.lastname: Morikawa
mesh_terms: D005656:Fungal Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D055646:Killer Factors, Yeast; D013058:Mass Spectrometry; D009183:Mycotoxins / Q000737:chemistry*; D011489:Protein Denaturation
chemical_list: D005656:Fungal Proteins; D055646:Killer Factors, Yeast; D009183:Mycotoxins