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MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME-OF-FLIGHT MASS SPECTROMETRY OF BACTERIAL RIBOSOMAL PROTEINS AND RIBOSOMES

SUH, MOO-JIN
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1106583486

Abstract Details

2005, PhD, University of Cincinnati, Arts and Sciences : Chemistry.
The ribosome is a ribonucleoprotein (RNP) complex that provides an ideal model for developing new analytical approaches for charactering RNPs. Here, I propose to develop suitable approaches for the systematic study of ribosomal proteins and ribosomes by mass spectrometry. As an initial trial, I have investigated the applicability of several common protein isolation procedures for their compatibility with downstream matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and numerated the Escherichia coli ribosomal proteins and their post-translational modifications with good sensitivity and high speed. This approach in combination with a ribosomal protein search algorithm was applied to Thermus thermophilus ribosomes. A major effort involved the use of limited proteolysis in combination with MALDI-MS to determine the stability of ribosomal proteins and to interrogate the interaction between ribosomal proteins and rRNA by monitoring the time-dependent production of proteolytic peptides and any remaining intact proteins. While exposed and unstructured proteins yield significant peptide ions via proteolytic digestion, proteins that are protected by rRNA in the inner part of the ribosome are not affected under the limited proteolysis conditions employed. Finally I expanded the use of limited proteolysis to the much larger and more complex 70S ribosomes from Thermus thermophilus to understand the role of ribosomal proteins in inter-subunit bridges. Extension regions of ribosomal proteins in inter-subunit bridges interact with rRNA and are important in maintaining the architecture of the complete 70S ribosome. It is envisioned that a combination of mass spectrometry and limited proteolysis can be used to further refine our understanding of the ribosome in various functional states, in particular, and also serve as a general method for characterizing other RNPs.
Dr. Patrick Limbach (Advisor)
175 p.
Mass Spectrometry, Matrix-Assisted Laser Desorption/Ionization, Ribosomes, RIbosomal Proteins, Post-translational Modification, Subcellular Proteomics, Limited Proteolysis;

Recommended Citations

Citations

  • SUH, M.-J. (2005). MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME-OF-FLIGHT MASS SPECTROMETRY OF BACTERIAL RIBOSOMAL PROTEINS AND RIBOSOMES [Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1106583486

    APA Style (7th edition)

  • SUH, MOO-JIN. MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME-OF-FLIGHT MASS SPECTROMETRY OF BACTERIAL RIBOSOMAL PROTEINS AND RIBOSOMES. 2005. University of Cincinnati, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1106583486.

    MLA Style (8th edition)

  • SUH, MOO-JIN. "MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME-OF-FLIGHT MASS SPECTROMETRY OF BACTERIAL RIBOSOMAL PROTEINS AND RIBOSOMES." Doctoral dissertation, University of Cincinnati, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1106583486

    Chicago Manual of Style (17th edition)

ucin1106583486
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© 2004, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.