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Date of Award
2-2014
Access Type
Campus Access
Document type
dissertation
Degree Name
Doctor of Philosophy (PhD)
Degree Program
Chemistry
First Advisor
Richard W. Vachet
Second Advisor
Igor A. Kaltashov
Third Advisor
Michael J. Knapp
Subject Categories
Analytical Chemistry | Chemistry
Abstract
This dissertation focuses on the mass spectrometric based methods for studying protein-metal binding. Identifying metal-protein interaction is a key step in understanding metal-binding protein structure and function. A phenomenon associated with gas phase dissociation behavior of metal-peptide complexes has been investigated. A positive correlation was found between the number of strong coordination groups in the peptide sequence and the degree of c and z ion formation after electron transfer dissociation of the peptide-metal complexes. Establishing thermochemical cycle enables a theoretical understanding of the process.
A new mass spectrometric method has been developed to identify Zn-bound His residues in Zn-metalloproteins relies on variations in the hydrogen deuterium exchange of the C2 hydrogen of His side chains. We show that this approach can be used to study the Zn-bound His residue in human &beta-2-microglobulin; (&beta2m;), a monomeric protein that has been shown to aggregate into amyloid fibrils in dialysis patients leading to dialysis-related amyloidosis.
The different effect of three divalent transition metals including Cu(II), Ni(II) and Zn(II) on &beta2m; oligomerization and fibril formation under physiological conditions is described. We found that Cu(II) can induced &beta2m; oligomerization and amyloidosis. In contrast, no oligomeric species can be formed with Ni(II), and only oligomers can be formed with Zn(II). A combination of metal catalyzed oxidation (MCO)-MS, hydrogen deuterium exchange (HDX)-MS and other spectroscopic techniques is utilized to obtain insights into the mechanism of Cu(II)-induced &beta2m; amyloidosis. We elucidate the different ways that these metals bind &beta2m;, thereby identifying key features of the &beta2m-Cu;(II) interaction that are essential for enabling this protein to form amyloid fibrils.
DOI
https://doi.org/10.7275/390r-fj04
Recommended Citation
Dong, Jia, "Mass Spectrometric Methods for Studying Protein-Metal Binding" (2014). Doctoral Dissertations 1896 - February 2014. 562.
https://doi.org/10.7275/390r-fj04
https://scholarworks.umass.edu/dissertations_1/562