Conformational studies of aqueous melittin: thermodynamic parameters of the monomer-tetramer self-association reaction

• Published in: Biochemistry (Easton) Vol. 22; no. 3; pp. 695 - 700
• Main Authors: Quay, Steven C, Condie, Claudia C
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.02.1983
• Subjects: Animals; Apidae; Apis mellifera; Bee Venoms; Bees; fluorescence; Hydrogen-Ion Concentration; Melitten; melittin; Osmolar Concentration; Protein Conformation; Solubility; Temperature; Thermodynamics; Water
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00272a026

The self-association reaction in which four melittin molecules associate to form an aqueously soluble tetramer was studied by fluorescent spectroscopy. At 23 degrees C, pH 7.15, gamma/2 0.50, the dissociation constant, Kd, is 3.20 x 10(-16) M3. At 23 degrees C, gamma/2 0.60, melittin has an amino acyl group with a proton ionization constant at ca. 10(-6) M, which must be un-ionized for tetramer formation to occur. The change in Kd with temperature indicates the forward reaction (tetramer formation) proceeds primarily by entropic changes, with delta H degrees = -20.3 kJ/mol of monomer and delta S degrees = 211 J/(K . mol of monomer). The observed enthalpic and entropic values for the tetramerization reaction are consistent with the expected contributions of both nascent hydrogen bonds and hydrophobic stabilization to the reaction. The ionic strength dependence of the tetramerization reaction was found to be consistent with an Edsall-Wyman treatment of activity coefficients. Specifically, the calculated charge of melittin varied from 2.5 (pH 10.53, gamma/2 less than 0.08) to ca. 6 (pH 7.15, gamma/2 greater than 0.3) and showed a strong dependence on gamma/2.