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Development and application of compa...

Zhao, Jia.

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Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry./
作者:	Zhao, Jia.
面頁冊數:	171 p.
附註:	Adviser: David M. Lubman.
Contained By:	Dissertation Abstracts International68-08B.
標題:	Chemistry, Analytical. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3276344
ISBN:	9780549181224

Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry.
Zhao, Jia.

Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry. - 171 p.

Adviser: David M. Lubman.

Thesis (Ph.D.)--University of Michigan, 2007.

Liquid phase glycoprotein enrichment, fractionation and glycoprotein microarray techniques based on lectin affinity, non-porous reverse phase HPLC (NP-RP-HPLC) and mass spectrometry characterization have been developed. These approaches provide improved sensitivity and throughput. Naturally occurring glycoproteins in human serum were screened and characterized for different glycan structures, allowing us to do comparative studies on monitoring individual glycoproteins in samples from different biological states. An application of this method in pancreatic cancer has revealed new potential marker glycoproteins. Both sialylation and fucosylation increased as a function of cancer for several proteins including Hemopexin, Kininogen-1, Antithrombin-III and Haptoglobin-related protein, whereas decreased sialylation was detected on plasma protease C1 inhibitor. Their alterations were further verified by western-blot and LC-MS experiments. In addition, the N83 glycosylation site of alpha1-antitrypsin was found to be down-regulated. A platform based on capillary hydrophilic interaction chromatography with on-line ESI-TOF MS for global N-glycans and glycosylation sites profiling in serum samples has been developed. 105 unique carbohydrate structures and 92 glycosylation sites were identified using this sensitive approach. 44 oligosaccharides were found to be distinct in pancreatic cancer serum, which might be used as a specific biomarker of malignancy. Their glycan structures were further elucidated by CID fragmentation using tandem mass spectrometry.

ISBN: 9780549181224Subjects--Topical Terms:

586156
Chemistry, Analytical.

Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry.
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245 1 0 $a Development and application of comparative proteomics and glycoproteomics approaches based on liquid phase separation and mass spectrometry.
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502 $a Thesis (Ph.D.)--University of Michigan, 2007.
520 $a Liquid phase glycoprotein enrichment, fractionation and glycoprotein microarray techniques based on lectin affinity, non-porous reverse phase HPLC (NP-RP-HPLC) and mass spectrometry characterization have been developed. These approaches provide improved sensitivity and throughput. Naturally occurring glycoproteins in human serum were screened and characterized for different glycan structures, allowing us to do comparative studies on monitoring individual glycoproteins in samples from different biological states. An application of this method in pancreatic cancer has revealed new potential marker glycoproteins. Both sialylation and fucosylation increased as a function of cancer for several proteins including Hemopexin, Kininogen-1, Antithrombin-III and Haptoglobin-related protein, whereas decreased sialylation was detected on plasma protease C1 inhibitor. Their alterations were further verified by western-blot and LC-MS experiments. In addition, the N83 glycosylation site of alpha1-antitrypsin was found to be down-regulated. A platform based on capillary hydrophilic interaction chromatography with on-line ESI-TOF MS for global N-glycans and glycosylation sites profiling in serum samples has been developed. 105 unique carbohydrate structures and 92 glycosylation sites were identified using this sensitive approach. 44 oligosaccharides were found to be distinct in pancreatic cancer serum, which might be used as a specific biomarker of malignancy. Their glycan structures were further elucidated by CID fragmentation using tandem mass spectrometry.
520 $a In another study, a protein profiling method based on chromatofocusing/NPS-RP-HPLC and online-ESI-TOF MS analysis was applied to study MCF10AT1 and the estrogen-treated MCF10AT1E2 cells. This method provides protein pI and intact MW information which are very useful for suggesting protein post-translational modifications and truncations. Key estrogen-regulated proteins were revealed. They were found to play an important role in signal pathways such as G1-S-specific cyclin and c-myc and have a role in the ras pathway such as GRB2-related adaptor protein, Mortalin. We have also utilized a 2-D liquid phase separation mass mapping technique to identify proteins differentially expressed in the progression from Barrett's metaplasia to esophageal adenocarcinoma. The changed protein expression was verified at the messenger level and further confirmed by immunohistochemistry, notably Rho GDP dissociation inhibitor 2 and alpha-enolase. These identified proteins may have utility as candidate markers of progression from Barrett's metaplasia to esophageal adenocarcinoma.
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