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Investigating copper binding to bacterial metallothioneins SmtA and MymT
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Tareen, Maria (2014) Investigating copper binding to bacterial metallothioneins SmtA and MymT. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b2751744~S1
Abstract
Metallothioneins are characterized as a group of small proteins with low molecular weight and high
content of cysteinyl residues and their synthesis is induced by metal ions (e.g., Zn or Cd). The physicochemical properties of Zn-, Cd-, and Cu-containing MTs have been studied. However, copper
metallothioneins (CuOMTs) have been examined less intensively, with only two MTs containing Cu(I)
having been structurally characterized to date i.e., yeast MT and Neurospora crassa MT.
The present study focuses on the stoichiometry and structural effects of binding of Cu(I) to the natively zinc binding MT (SmtA) from the cyanobacterium Synechococcus elongatus PCC7942. For
this purpose, a Cu(I) titration study of Zn4SmtA was anaerobically performed utilizing the collective
detection of ESIOMS, UV-Vis absorbance and 1H-15N NMR spectroscopy, and fully exchanged Cu(I)-SmtA was also generated from the apoOprotein. ESIOMS data confirmed the presence of predominantly monomeric Cu7SmtA. UV-vis titrations of Zn4SmtA showed that up to 7 Cu(I) displace Zn(II) from the protein, which was further studied in more detail by Cu(I) titrations of Zn4SmtA monitored with 1H-15N and 111Cd-15H NMR spectroscopy. The extensive NMR analysis showed the unfolding of protein on Cu(I) addition with absence of inertness towards Cu(I) exchange and that the protein undergoes significant conformational changes on Cu(I) addition.
Preliminary characterization of MymT from Mycobacterium tuberculosis was also done and the composition of the metal cluster in the dominant Cu5MymT species was proposed and cooperativity
was established by ESIOMS analyses, although structural characterisation was impeded by the highly
dynamic nature of the protein.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Library of Congress Subject Headings (LCSH): | Metallothionein, Copper | ||||
Official Date: | April 2014 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | Department of Chemistry | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Blindauer, Claudia A. | ||||
Extent: | xxii, 190 leaves : illustrations, charts | ||||
Language: | eng |
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