Graduate Thesis Or Dissertation
 

Physical studies of the yeast inner histones

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  • In this work, the four yeast inner histones were prepared by the use of consecutive slab gel fractionation. The use of this technique allowed the first complete fractionation and rigorous identification of the four yeast inner histones, and made possible a study of their complexing behavior. Variants and adjuncts of preparative electrophoresis by the method of gel slicing are presented. These include improved methods of locating and eluting protein bands. The yeast histones were located in the gel using the phosphorescence detection technique of Isenberg et al. (1975). Elution was achieved conveniently and efficiently using an all-glass gel eluter of original design. The method permits the separation of closely spaced components and may be used in whole, or in part, to isolate a wide variety of proteins in pure form from crude extracts. The complexing pattern of the yeast inner histones was investigated using fluorescence anisotropy and circular dichroism. The pattern was found to be the same as that found previously for calf and pea histones. In particular, strong interactions were observed for the following pairs of yeast histones: H2a-H2b, H2b-H4, and H3-H4. Interspecies binding constants for calf and yeast histones were measured and found to be close to the intraspecies binding constants. This result indicates a high degree of conservation of binding surfaces.
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