[Protein composition of DNA-matrix complexes with "strong" and "weak" types of bonds, isolated from Ehrlich ascites
Abstract:
A preparative procedure for obtaining tightly and loosely bound DNA-matrix complexes was developed. To this end, the DNA-matrix complexes were separated from DNA excess by restrictase digestion and solubilized in 0.1% sodium dodecyl sulfate + 8 M urea. To obtain tightly bound complexes, the lysate was transferred to a solution containing 2 M LiCl and 4 mM urea and gel filtered in the same medium. Tightly bound complexes were eluted in the same peak with DNA. Loosely bound complexes were obtained by gel filtration in 0.1% sodium dodecyl sulfate + 8 M urea after preliminary digestion of the original complexes with DNAase I which selectively destroys the tight bonds. Within the composition of loosely bound DNA-matrix complexes, the polypeptides with Mr of 180, 65-75, 58 and 47-50 kDa were identified, whereas tightly bound complexes were shown to contain polypeptides with molecular masses of 180, 65-75, 63, 61, 58 and 52-53 kDa.
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