Histone variants and post-translational modifications (PTMs) are known to play central roles in genome regulation and maintenance. Quantitavely characterization of the variants and the associated PTMs is the first step to understand their biological functions. However, many variants and the PTM combinations are inaccessible by antibody-based methods or bottom-up tandem mass spectrometry. For many, the only tractable approach is with intact protein top-down tandem mass spectrometry. In this dissertation, a method of identification of histone H4 proteoforms is developed, revealing the challenge of top-down proteomics. Then a method of proteoform identification and relative quantitation of histones H2A and H2B by FT-ICR MS and MS/MS is developed, yielding quantitative identification of all detected H2A and H2B isobaric and isomeric proteoforms with a label-free approach in HeLa. The method is further extended to 1) normal and HIV-infected U937 cells; 2) the Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) stem cells sorted into early G1, late G1, S, and G2/M phases; 3) MDA-MB-231 breast cancer cells synchronized to S and M phases. The top-down MS/MS approach provides a path forward for more extensive elucidation of the biological roles of many previously unstudied histone variants and PTMs. / A Dissertation submitted to the Department of Chemistry and Biochemistry in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Summer Semester 2016. / June 27, 2016. / Includes bibliographical references. / Alan G. Marshall, Professor Directing Dissertation; David M. Gilbert, University Representative; Michael G. Roper, Committee Member; Timothy M. Logan, Committee Member.
| Identifer | oai:union.ndltd.org:fsu.edu/oai:fsu.digital.flvc.org:fsu_366435 |
| Contributors | Dang, Xibei (authoraut), Marshall, Alan G. (Alan George), 1944- (professor directing dissertation), Gilbert, David M. (university representative), Roper, Michael Gabriel (committee member), Logan, Timothy M., 1961- (committee member), Florida State University (degree granting institution), College of Arts and Sciences (degree granting college), Department of Chemistry and Biochemistry (degree granting department) |
| Publisher | Florida State University, Florida State University |
| Source Sets | Florida State University |
| Language | English, English |
| Detected Language | English |
| Type | Text, text |
| Format | 1 online resource (111 pages), computer, application/pdf |
| Rights | This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). The copyright in theses and dissertations completed at Florida State University is held by the students who author them. |
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