RESUMO
Micromethods for measurements of electric conductivity, transference numbers and concentrations of inorganic ions within immobilized protein crystals have been developed and applied to study tetragonal lysozyme crystals cross-linked with glutaraldehyde. Donnan equilibria and mobilities of ions in this crystal were calculated using the data of these methods and the data of crystal pH titration. Taken together these results characterize the lysozyme crystal as an ion exchanger whose electrical properties and ion composition differ greatly from those of the external solution. Although anions transfer most of the current in the crystals, anion mobility is considerably lower than that of cations. Mobility of all ions in the crystal is considerably lower than in solution (3.5-50 times for cations and 120-330 times for anions) and depends on steric restrictions and charges of both ions and lysozyme molecules. Similar features in behavior of crystalline and biological channels are discussed.
Assuntos
Muramidase/química , Fenômenos Químicos , Físico-Química , Reagentes de Ligações Cruzadas/química , Cristalização , Condutividade Elétrica , Ativação Enzimática , Glutaral/química , Concentração de Íons de Hidrogênio , Canais Iônicos/química , Íons , Concentração Osmolar , Cloreto de Sódio/químicaRESUMO
Diffusion of synthetic dyes in lysozyme crystals and bovine serum albumin (BSA) amorphous films is shown to depend on the size of diffusate molecule far more strongly than in solutions. The diffusion coefficient increases from 10(-8) to 10(-13) cm2 sec-1 on about 1.5 divided by 2--fold increase in diffusate dimensions, the diffusion being completely arrested on reaching the limiting dimensions of about 14 A and 12 A for BSA films and lysozyme crystals, respectively. The diffusion in lysozyme crystals is anisotropic, with a diffusion maximum directed along their screw axes. The diffusion coefficient dependence on the charge of diffusate and its binding to proteins is discussed.
