Thesis
The folding mechanisms of the eukaryotic and archaeal histones
Washington State University
Master of Science (MS), Washington State University
2012
Handle:
https://hdl.handle.net/2376/102496
Abstract
The focus of this thesis is to study the folding mechanism of two histone proteins, the eukaryotic histone heterodimer H2A-H2B and the archaeal histone homodimer hMfB. These proteins have similar structure and function, but share only 25% similarity in sequence and have different populated intermediates: H2A-H2B populates a monomeric and a dimeric intermediate and hMfB populates a monomeric intermediate. A FRET system was developed to study the dimerization reaction of H2A-H2B. Equilibrium experiments showed that an engineered FRET system only modestly destabilizes H2A-H2B. Fluorescence anisotropy confirmed that the fluorophores are not buried. Equilibrium and kinetic studies were performed on twelve hMfB mutants. The mutations were designed to probe specific residues and determine their role in the development of structure in the dimerization transition state. Unfolding kinetic data indicate that residues in the central and C-terminal helices stabilize contribute significantly to the stability of the monomeric intermediate and the transition state. Folding kinetic data indicate that native-like structure is common to the transient species throughout the folding reaction of hMfB, which is in contrast to H2A-H2B, whose intermediates have significant non-native structure.
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Details
- Title
- The folding mechanisms of the eukaryotic and archaeal histones
- Creators
- Anna M. Lindsay
- Contributors
- Lisa M. Gloss (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Molecular Biosciences, School of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525019901842
- Language
- English
- Resource Type
- Thesis