Pressure-induced dissociation and denaturation of allophycocyanin at subzero temperatures

The thermodynamics of assembly of the allophycocyanin hexamer was examined employing hydrostatic pressures in the range of 1 bar to 2.4 kbar and temperatures of 20 to -12 degrees C, the latter made possible by the decrease of the freezing point of water under pressure. The existence of two processes, dissociation of the hexamer into dimers, (alpha beta)3-->3 (alpha beta), and dissociation of the alpha beta dimers into monomers, (alpha beta)-->alpha + beta have been recognized previously by changes in the absorbance and fluorescence of the tetrapyrrolic chromophores owing to added ligands. The same changes are observed in the absence of ligands at pressures of under 2.4 kbar and temperatures down to -12 degrees C. On decompression from 2.4 kbar at 0 degrees C, appreciable hysteresis and a persistent loss of 50% in the absorbance at 653 nm is observed. It results from the conformational drift of the isolated subunits and is reduced to 10% when the highest pressure is limited to 1.6 kbar. The thermodynamic parameters of the reaction alpha + beta-->alpha beta can be determined from pressure effects on perchlorate solutions of allophycocyanin, which consist of dimers alone. Their previous knowledge permits estimation, under suitable hypotheses, of the thermodynamic parameters of the reaction 3(alpha beta)-->(alpha beta)3 from the overall pressure effects on the hexamers. Both association reactions have positive enthalpy changes, and the whole hexamer assembly is made possible by the excess entropy.