Dissertation
Calsequestrin
Washington State University
Doctor of Philosophy (PhD), Washington State University
01/2016
DOI:
https://doi.org/10.7273/000002430
Handle:
https://hdl.handle.net/2376/116924
Abstract
We studied the structure-function relations and biophysical characteristics of skeletal calsequestrin by determining the Ca2+-complex crystal structures of human (H. sapiens sapiens) and rabbits (O. cuniculus) skeletal calsequestrins. The Ca2+-complex structures showed calsequestrin binds Ca2+ in distinct sites, and from them, we proposed a structure-based Ca2+-dependent polymerization model. We then characterized two human skeletal calsequestrin mutants implicated in malignant hyperthermia (M53T mutant) and vacuolar aggregates myopathy (D210G mutant) and found that the M53T mutation eliminates a hydrophobic interaction important for dimerization, while the D210G mutation destabilizes a structural Ca2+-binding site, making D210G the first known disease-associated mutation to a single Ca2+ binding site. Finally, to see if calsequestrin’s characteristics were as conserved across mammalian life as their amino acid sequences implied, we characterized the post-translational modifications to cardiac (glycosylation and phosphorylation) and skeletal calsequestrin (glycosylation only) in humans (only cardiac), cattle (B. taurus), mice (M. musculus), and rats (R. norvegicus). Using the low- and high-Ca2+ forms of native and recombinant skeletal calsequestrin from cattle, we showed glycosylation’s effect on calsequestrin’s Ca2+-dependent structural changes and polymerization. We compared those observations to bottlenose dolphins (T. truncatus), whose skeletal calsequestrin is almost identical to cattle’s, despite their different environments and physiology.
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Details
- Title
- Calsequestrin
- Creators
- Kevin Michael Lewis
- Contributors
- ChulHee Kang (Advisor)James A Brozik (Committee Member)Jeffrey P Jones (Committee Member)Kirk A Peterson (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Chemistry, Department of
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Publisher
- Washington State University
- Number of pages
- 123
- Identifiers
- 99900581833301842
- Language
- English
- Resource Type
- Dissertation