Derivatization of peptides for structural studies by mass spectrometry /
Mass spectrometric sequencing of arginine-containing peptides is improved by selective derivatization of the guanidino groups with acetylacetone. The significant increase in the relative abundance of sequence specific fragment ions is observed in matrix assisted laser desorption/ionization post-sou...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
2001.
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| Subjects: | |
| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=728908911&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | Mass spectrometric sequencing of arginine-containing peptides is improved by selective derivatization of the guanidino groups with acetylacetone. The significant increase in the relative abundance of sequence specific fragment ions is observed in matrix assisted laser desorption/ionization post-source decay (MALDI-PSD) mass spectra. The fragmentation efficiency increases by a factor of 2-3.5 with the best results for peptides with derivatized residue at the N-terminus or both ends of the sequence. This strategy makes it possible to obtain useful structural information from arginine-containing peptides that otherwise do not fragment well. Three new heterobifunctional photoreactive cross-linkers are designed and synthesized to study peptide-protein and peptide-peptide interactions by MALDI. The cross-linkers contain a thiol-reactive maleimide group, a photoreactive benzophenone group and a cysteic acid residue. The cross-linkers differ in the length of the spacer arm that has one, two or five methylene groups in it. Fragmentation pathways of the cross-linkers and three cross-linker-peptide conjugates are studied by MALDI-PSD in negative and positive ion mode. Ribonuclease S-peptide analog is successfully cross-linked to S-protein. The cross-linking is unambiguously confirmed by MALDI, SDS-PAGE and HPLC. The other investigated systems include leucine zipper peptides and an analog of bumblebee venom peptide bombolitin III. The latter is studied by nanoelectrospray ionization (nano-ESI) mass spectrometry. |
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| Item Description: | Vita. "Major Subject: Chemistry". |
| Physical Description: | xiv, 147 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilm Inc. |
| Bibliography: | Includes bibliographical references (leaves 135-145). |