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Structure and properties of rat-thymus deoxyribonucleoprotein. A comparison of the electric birefringence and other properties of deoxyribonucleoprotein prepared by various methods.

ABSTRACT: 1. A comparison has been made of the properties of dNP (deoxyribonucleoprotein) prepared in various ways from rat thymus. The variables studied were: (a) extent of homogenization; (b) concentration of various metal ions; (c) concentration of EDTA; (d) extent of dialysis; (e) concentration of dNP. The chief method used for studying the dNP was that of electric birefringence; this gives information about the optical and electrical properties of molecules oriented in an electric field and about their length. 2. The extent of homogenization during the preparation affects the birefringence properties only slightly and the solubility properties of the constituent DNA not at all. However, the protein content of the dNP decreases after repeated homogenization. 3. In the presence of added Ca(2+), Mg(2+) or Na(+) an insoluble product is obtained. 4. The presence of added EDTA affects the birefringence properties only slightly. 5. In the final stage of preparation, both very limited dialysis and extensive dialysis (in terms of volume ratio) cause partial insolubility of the final product. 6. dNP prepared at low concentrations resembles in some respects dNP prepared at higher concentrations and then subjected to limited enzyme action but it is not degraded (as by extensive enzyme action); ultracentrifuge studies of the constituent DNA confirm that no degradation of the DNA has occurred. 7. It is concluded that dNP prepared at low concentrations or by extensive dialysis is denatured and that preparation of ;native' material is favoured by maintenance of a high dNP concentration and by fairly limited dialysis.

SUBMITTER: Itzhaki RF

PROVIDER: S-EPMC1265112 | biostudies-other | 1966 Jul

REPOSITORIES: biostudies-other

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Structure and properties of rat thymus deoxyribonucleoprotein. Solubility properties and effects of precipitation.

Project description:1. Deoxyribonucleoprotein was prepared from rat thymus and was studied chiefly by the method of electric birefringence. The birefringence depends on the electrical and optical properties of the molecules and on their volume; the decay time of birefringence (after the removal of the electric field) depends on molecular length. 2. A comparison was made of the properties of deoxyribonucleoprotein redissolved after precipitation in 0.15m-sodium chloride with those of the original material, the main object being to find if structural changes have occurred in the former. As a preliminary, the dependence of the solubility of the deoxyribonucleoprotein on the concentration of added salt was investigated, and the birefringence properties were also studied after the addition of sodium chloride at low concentrations, after the alteration of pH and after dialysis. 3. It was found that precipitation of deoxyribonucleoprotein occurs in two fractions, beginning at about 0.4mm-sodium chloride. The solubility is minimal at about 0.15m-sodium chloride. 4. The electric birefringence and decay time of the deoxyribonucleoprotein decrease with increasing concentration of added sodium chloride, and the birefringence also decreases after dialysis. Prolonged dialysis leads to precipitation. 5. The properties of deoxyribonucleoprotein redissolved after precipitation with 0.15m-sodium chloride differ considerably from those of the original deoxyribonucleoprotein. This is attributed to some type of disorganization process occurring during precipitation. It is concluded that the organization of the original structure is very specific.

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