Sedimentation-equilibrium and other physicochemical studies on the lysine-rich fraction of calf thymus histones

Abstract

1. The lysine-rich fraction (Ia+Ib, or f1) of calf thymus histones was isolated as the sulphate by acid extraction. 2. Sedimentation-equilibrium measurements with interference optics showed that this fraction was monodisperse with a molecular weight of 19500±2000. 3. The ‘apparent molecular weight’ calculated from the sedimentation-equilibrium studies varied markedly with concentration. The large second virial coefficient implied by such variation was attributed to the very high charge/mass ratio of this relatively small protein. Estimates of the charge were made from the values of this virial coefficient. 4. The very large value of the virial coefficient explains anomalies in the earlier reports of the molecular weight of this histone and also why the z-average molecular weight can appear to be lower than the weight-average molecular weight. 5. The differences of the specific refractive increments, and the partial specific volumes, between dialysed and undialysed solutions of this histone fraction could also be attributed to its high molecular charge, which was estimated from these differences and agreed, within the expected limits, with the value deduced from the second virial coefficient. 6. Sedimentation-velocity measurements combined with the known molecular weight imply that lysine-rich histone has a high frictional ratio and an extended shape. Optical-rotatory-dispersion measurements indicated that it had a low helical content.