Characterization of the dissociation equilibria of the histone H3H4 tetramer using sedimentation velocity analytical ultracentrifugation
Date
2020
Authors
Connolly, Mark Edward, author
Hansen, Jeffrey, advisor
Nyborg, Jennifer, committee member
Walrond, John, committee member
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Abstract
By performing sedimentation velocity analytical ultracentrifugation experiments under various ionic and pH conditions the kd can be measured. This system was found to be pH dependent with a change of Svedberg (S) value distribution in between pH 5.2 and pH 5.5. The H3H4 system was found to be an interacting one due to the change in S value distribution with increasing the histone protein concentration. We found that the S value distribution is highly dependent on the ionic conditions of the solution with 2M NaCl solution showing higher S values then 5mM KPO4 solution at the same concentration and pH. Oddly enough adding HEPES to a KPO4 buffer will destabilize the H3H4 species present with 5mM KPO4 having a higher S value distribution than 5mM KPO4 10mM HEPES at the same concentration and pH. I was unable to model these systems to a H3H4 dimer to tetramer equilibrium model which leads me to believe the conditions I was using did not stabilize the tetramer save for the 2M NaCl.
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Subject
dimer
H3H4
tetramer
dissociation
AUC
histone