Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List


Thesis_final_v6_DI.pdf (4.79 MB)

ETD Abstract Container

Abstract Header

Elucidating the Sequence and Structural Features of Human Bence-Jones Proteins

Perera, Weliwaththage Thilini
http://rave.ohiolink.edu/etdc/view?acc_num=toledo1526078353370352

Abstract Details

2018, Master of Science, University of Toledo, Chemistry.
Amyloidosis diseases are characterized by the deposition of insoluble protein aggregates called amyloids. More than 30 diseases are associated with amyloid-forming proteins, including Alzheimer’s, Parkinson’s, Huntington’s and immunoglobulin light chain amyloidosis (AL amyloidosis).The proteins contributing to each disease have distinct primary structures. The mechanism of amyloid formation is not well understood, but appears to be associated with protein misfolding processes accompanied by self-aggregation. In AL amyloidosis, immunoglobulin light chain proteins become misfolded and accumulate as amyloid. No effective therapeutic solution is available to treat this disease condition, which is usually fatal a few months after diagnosis. AL amyloidosis is a subset of patients with multiple myeloma (MM), a malignant disease condition characterized by bone marrow failure. Most of the patients having MM excrete monoclonal free immunoglobulin light chains, also called Bence-Jones proteins, into the urine. In contrast to AL amyloidosis, amyloid deposits are not observed in vivo in most patients suffering from MM. Consequently, the identification of the precise sequence and structural information of proteins related to MM are essential to understand the factors responsible for aggregation. As reported in the literature, several analytical methods have been used to study primary, secondary, tertiary and quaternary structures of amyloid-forming proteins. In our study, mass spectrometry, circular dichroism (CD) spectroscopy and dynamic light scattering (DLS) were used to obtain sequence and structural information about immunoglobulin light chain proteins, isolated from urine of AL amlylodosis and MM patients. Accordingly, molecular masses of seven Bence-Jones protein samples were measured by ESI-MS and MALDI-MS and the properties of their ions in the gas phase were observed by IMS-MS. Since five of them are with unknown sequences, the bottom-up proteomic approach was used to obtain unreported sequences of immunoglobulin light chain proteins. The enzyme-digested protein samples were analyzed by MALDI-MS, ESI-MS, IMS-MS and HPLC-ESI-MS/MS. A de novo sequencing assisted-data base search was performed using PEAKS search tool to obtain possible peptide sequences.Multistep MS-based approach was used to differentiate between leucine and isoleucine residues present in newly identified peptides. Furthermore, CD spectroscopy was used to compare the secondary structure elements of different light chain proteins. Most light chains showed high percentage of ß-sheets, which is common for amyloid-forming proteins. In addition, DLS was used to study the effect of physiochemical parameters on the aggregation behavior of light chain proteins. Overall, novel sequence and structural features of immunoglobulin light chains were obtained by these approaches and will be correlated with the properties of other amyloid-forming proteins.
Dragan Isailovic (Advisor)
Leif Hanson (Committee Member)
John Bellizzi (Committee Member)
Chemistry

Recommended Citations

Citations

  • Perera, W. T. (2018). Elucidating the Sequence and Structural Features of Human Bence-Jones Proteins [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1526078353370352

    APA Style (7th edition)

  • Perera, Weliwaththage Thilini. Elucidating the Sequence and Structural Features of Human Bence-Jones Proteins. 2018. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=toledo1526078353370352.

    MLA Style (8th edition)

  • Perera, Weliwaththage Thilini. "Elucidating the Sequence and Structural Features of Human Bence-Jones Proteins." Master's thesis, University of Toledo, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1526078353370352

    Chicago Manual of Style (17th edition)

toledo1526078353370352
271
© , some rights reserved.Creative Commons License Elucidating the Sequence and Structural Features of Human Bence-Jones Proteins by Weliwaththage Thilini Perera is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by University of Toledo and OhioLINK.