Reversible Dissociation of Crystalline Yeast Phosphoglyceric Acid Mutase in Alkali

Abstract

The structure of crystalline yeast phosphoglyceric acid mutase has been investigated by sedimentation-velocity and equilibrium measurements, optical rotatory dispersion measurements and viscometry. The data indicate that this enzyme is a globular, compact and highly organized protein with a low helix content. The native structure remains unchanged at pH 10.5. Dissociation of the enzyme into subunits has been observed at pH values of 11.5 and above. From optical rotatory dispersion measurements, it is found that the enzyme loses a large part of its organized conformation when it dissociates in alkaline solution. On neutralization, the alkali-treated enzyme regains its activity. The ability to regain the enzyme activity is gradually lowered with the increase of pH value to be incubated and with time of exposure. Inactivation at pH 13.0 is almost irreversible. However, the reversibility of the inactivation at pH 13.0 is appreciably enhanced by the presence of phosphate compounds in the reactivation system. Particulary, it is found that presence of substrates or the coenzyme is effective for considerable improvement of the reversibility. Molecular weight analyses by ultracentrifugation indicate that subunits have approximately equal molecular weights and that the native enzyme is consisted of four polypeptide chains.