The charge structure and ionic interactions of elastin prepared from the pig thoracic aorta by acid, alkali, or CNBr extraction have been investigated by potentiometric titration and radiotracer techniques. The number of charged groups was consistent with the amino acid composition, comparable to elastin from other sources and insensitive to the method of preparation. The enthalpies of ionization of the basic groups were comparable for those previously found for proteins but those of the acidic groups were higher. Ionic interactions were predominantly electrostatic although a strong affinity for chloride ions was noted. Changes in ionic interactions as the elastin was stretched had a similar effect to an increase in the apparent fixed charge density of the tissue. Mechanical strain altered the protonation of the elastin and the pK of the carboxyl groups. Conversely, the conformation of the elastin network varied with ionic strength and pH, being particularly sensitive to the degree of ionization of the more basic groups and with the ionic strength and anion composition of the medium. We speculate that strain induced changes in the conformation of elastin altering its reactivity towards lipids, ions or matrix macromolecules or changes in its mechanical properties resulting from changes in its ionic environment may be of physiological or pathological importance.
Radiotracer methods have been used to measure the distributions of inorganic ions in the aorta. Data on fresh tissue were generally consistent with previous measurements and showed UCa greater than UI greater than UNa. In order to relate these measurements to the macromolecular composition of the arterial wall, the effects of various interventions--cell lysis, autoclaving, and NaOH digestion--were investigated. Elastin was found to be the major determinant of ionic distribution coefficients, to possess a slight positive charge under physiological conditions, and to display a high affinity for Ca++. The distribution coefficients did not conform to the predictions of the Donnan theory of ionic equilibrium and the degree of nonideality, as measured by the fixed charge density and the ion selectivity coefficients, varied with ionic strength. Data are also presented on the distribution of neutral solutes of different molecular weights in the various arterial preparation, in order to provide a basis for comparison of the behaviour of charged solutes.
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