Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry
碩士 === 國立暨南國際大學 === 應用化學系 === 100 === Reversible oxidative cysteine modifications play a critical role in the redox-based signaling mechanism. Dynamic profiling oxidation state of the cysteine residues in protein such as Peroxiredoxin (Prx) presents a formidable challenge. Here, we present a novel a...
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ndltd-TW-100NCNU05000212015-10-13T21:12:10Z http://ndltd.ncl.edu.tw/handle/37663862666571767886 Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry 結合二氧化鈦奈米鑽石萃取技術及質譜分析技術於蛋白質半胱胺酸氧化狀態的分析 Chen, Chun-Hua 陳俊樺 碩士 國立暨南國際大學 應用化學系 100 Reversible oxidative cysteine modifications play a critical role in the redox-based signaling mechanism. Dynamic profiling oxidation state of the cysteine residues in protein such as Peroxiredoxin (Prx) presents a formidable challenge. Here, we present a novel approach to selectively isolate sulfopeptides (peptides containing cysteine sulfonic acid and sulfinic acid) from complex digests using TiO2-coated nanodiamonds (TiO2-coated NDs) prior to MS analysis. The method was applied to selectively concentrate sulfopeptides from either a highly dilute solution or a highly complex peptide mixture. This method allowed us to identify the 22 distinct cysteine oxidation status out of a total 35 present in performic-acid-oxidized BSA by MALDI-TOF MS and all the distinct cysteine oxidation status by LC-ESI-MS/MS. Finally, we applied the new approach to identify the cysteine oxidation status of hydrogen peroxide–treated Prx and BSA by LC-ESI-MS/MS. Cysteine residues were found to display in either cysteine sulfonic acid or cysteine sulfinic acid status after hydrogen peroxide treatment. Enhanced detection of low abundance sulfopeptides containing active cysteine positions (C52, C71, C83, and C173) was achieved due to the highly selective enrichment using TiO2-coate NDs. Wu, Chih-Che 吳志哲 2013 學位論文 ; thesis 56 zh-TW |
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碩士 === 國立暨南國際大學 === 應用化學系 === 100 === Reversible oxidative cysteine modifications play a critical role in the redox-based signaling mechanism. Dynamic profiling oxidation state of the cysteine residues in protein such as Peroxiredoxin (Prx) presents a formidable challenge. Here, we present a novel approach to selectively isolate sulfopeptides (peptides containing cysteine sulfonic acid and sulfinic acid) from complex digests using TiO2-coated nanodiamonds (TiO2-coated NDs) prior to MS analysis. The method was applied to selectively concentrate sulfopeptides from either a highly dilute solution or a highly complex peptide mixture. This method allowed us to identify the 22 distinct cysteine oxidation status out of a total 35 present in performic-acid-oxidized BSA by MALDI-TOF MS and all the distinct cysteine oxidation status by LC-ESI-MS/MS. Finally, we applied the new approach to identify the cysteine oxidation status of hydrogen peroxide–treated Prx and BSA by LC-ESI-MS/MS. Cysteine residues were found to display in either cysteine sulfonic acid or cysteine sulfinic acid status after hydrogen peroxide treatment. Enhanced detection of low abundance sulfopeptides containing active cysteine positions (C52, C71, C83, and C173) was achieved due to the highly selective enrichment using TiO2-coate NDs.
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author2 |
Wu, Chih-Che |
author_facet |
Wu, Chih-Che Chen, Chun-Hua 陳俊樺 |
author |
Chen, Chun-Hua 陳俊樺 |
spellingShingle |
Chen, Chun-Hua 陳俊樺 Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
author_sort |
Chen, Chun-Hua |
title |
Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
title_short |
Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
title_full |
Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
title_fullStr |
Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
title_full_unstemmed |
Selective Profiling of Protein Cysteine Oxidative Modifications by TiO2 Enrichment and Mass spectrometry |
title_sort |
selective profiling of protein cysteine oxidative modifications by tio2 enrichment and mass spectrometry |
publishDate |
2013 |
url |
http://ndltd.ncl.edu.tw/handle/37663862666571767886 |
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