Calexcitin B: a new calcium-sensor protein in the nervous system of the squid
By: by Gombos, Zoltan.
Published: 2004
Language: eng
Number of pages: 171
ISBN-13: 9780494159859
Description: Calcium ions (Ca 2+ ) are paramount for cell viability and function. The Ca 2+ signalling pathways utilize specialized proteins, called Ca 2+ binding proteins (CaBPs). Calexcitin (CE) is a CaBP that putatively acts as an intracellular Ca 2+ -mediated messenger. CE is expressed specifically in the nervous system, and may play a critical role in memory. In this work, I elucidated the correct primary structure of CE and characterized its biochemical and biophysical properties. CE can bind three Ca 2+ at affinities similar to other Ca 2+ -sensor proteins. In addition, I have shown that CE is a member of the sarcoplasmic Ca 2+ binding protein (SCP) subfamily of the EF-hand superfamily. Like other SCPs, CE can bind Mg 2+ and has a complex Ca 2+ /Mg 2+ interplay that affects its structure. My studies have identified the physiological role of Mg 2+ binding and has shown that CE is a two domain protein that exists in three distinct conformational states: Apo-CE, has molten-globule characteristics; Mg 2+ -loaded CE possesses one native domain and the other domain is molten-globule like; Ca 2+ -loaded CE has both domains in native configuration. This indicates that under physiological concentrations of Mg 2+ and Ca 2+ , Mg 2+ -CE represents the resting state of the molecule and Ca 2+ -CE is the active state. The backbone nuclei of CE has been assigned by nuclear magnetic resonance and the results show that CE is composed of nine a-helices, eight of which form the four EF-hands and the ninth is near the C-terminus. This result clarified previous discrepancies in the literature regarding the secondary structure of CE and lays the foundation for studying the protein dynamics of CE. Models of CE, based on related proteins, show that, in agreement with biophysical data, CE is composed of two domains that do not interact considerably. Furthermore, in agreement with biophysical data, the model shows a putative recessed hydrophobic pocket that may be involved in target recognition, analogous to other Ca 2+ -sensor proteins. This thesis provides the basis for future investigations into elucidating the precise physiological function of CE. In addition, it enhances our understanding on how CaBPs work and how we can exploit their function for practical neuroprotective applications.
Notes: Thesis (Ph.D.) -- University of Toronto, 2004.

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