Conformational changes in histone IV as induced by salts and nucleoside triphosphates Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/zk51vk13p

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  • The addition of salts or nucleoside triphosphates to histone IV solutions induces conformational changes in the histone molecule. These conformational changes are investigated by measurement of the fluorescence anisotropy of the histone tyrosine emission, the circular dichroism (CD) of the peptide bond and of the tyrosine CD. Changes induced by Na₂HPO₄, NaH₂PO₄, Na₂SO₄, NaCl0₄, NaCl, MgCl₂, NaF, ADP, ATP, GTP, CTP and. TTP are investigated. Salt induced conformational changes are separated into two steps which occur on vastly different time scales. There is a change which is very fast and recorded simply as a rise. At sufficiently high salt concentration the fast change is followed by a slower one which takes minutes to hours depending on solution conditions. CD results demonstrate that a-helix formation accompanies the fast step. Fluorescence anisotropy measurements indicate that other conformational folding, to which the CD is insensitive, and histone dimerization also occur in the fast step. Equations are developed for a quantitative comparison of the salts studied. The slow step is found to be highly co-operative having a sharp onset at a given salt concentration followed by a steep rise to a maximum. CD spectra and kinetic CD measurements demonstrate that the slow step involves intermolecular β-sheet formation. Both static and kinetic measurements indicate that there is more than one process occurring in the slow step. Nucleoside triphosphates induce conformational changes at nucleotide concentrations which are of the same order of magnitude as the histone concentration. ATP and GTP interact with histone IV so strongly that plots of fractional conformational change versus nucleotide/histone are indistinguishable over a four fold range of histone concentrations. CTP and TTP are less effective but are at least two orders of magnitude more effective than any of the inorganic salts studied.
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