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Identification of protein acetylation

Author: ZhuYePing
Tutor: YangPiYuan
School: Fudan University
Course: Analytical Chemistry
Keywords: Post-translational modification Acetylated protein Mass Immunoprecipitation
CLC: Q51
Type: Master's thesis
Year: 2008
Downloads: 368
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Abstract

Acetylation is a reversible and highly regulated post-translational modification of proteins, mainly in the protein lysine residue ε-NH ₂ bit, which regulate apoptosis, subcellular localization, DNA and protein-protein interactions, DNA replication and repair, DNA transcription activity and stability of the protein and other protein-related processes. Lysine acetylation and regulation of enzymes and life of its occurrence aging process and some major diseases such as cancer, cardiovascular disease, there is a close link, thus protein acetylation identification of post-translational modifications have important biological significance. Traditional acetylated protein identification methods are immunoaffinity radioactive detection and detection, but these methods can only detect the presence of protein acetylation, can get specific information acetylation sites. Soft ionization mass spectrometry ESI-MS technique and MALDI-MS can be well applied to the identification of post-translational modification, can not only detect the protein post-translational modification, and can obtain detailed information modification sites. This thesis work aims to investigate two types of bio-mass spectrometry ESI-MS and MALDI-MS in the post-translational modification of protein acetylation Identification and Its Application in mouse liver acetylation identification of spectrum. The first chapter of this thesis on post-translational modifications in proteomics research and development, the identification of a variety of post-translational modification research strategy, post-translational modification acetylation research are outlined. The research work is mainly divided into two parts, namely, in the second and third chapters in introduced in detail. Chapter II of the study, the use of two types of bio-mass spectrometry ESI-MS (LC-IT-TOF mass spectrometer, LTQ-Orbitrap mass spectrometer) and MALDI-MS (4700 Proteomics Analyzer mass, QIT mass spectrometer), the acetylation acetylated BSA standard proteins identified by mass spectrometry studies of mouse liver acetylation Identification of protein modifications spectra provide methodological reference. Experiments show that, LC-ESI-MS (LTQ-Orbitrap) and MALDI-TOF-TOF (4700 proteomic analyzer) are two ways to identify acetylated protein mass than the other two methods is superior, and the two methods identified in the acetylation modification having complementary properties. In the third chapter of the study, we designed according to the second chapter of the two mouse liver acetylated protein identification line of research, one for the use of the mature protein separation technology two-dimensional gel electrophoresis (2DE) isolated rat liver whole protein, using the teacher Zhao Shiming Biomedical Research Institute provided lysine acetylation antibody 2D Western-Blotting validation, Western-Blotting Western blot analysis were positive points compared to 2DE gels and screened agarose gel digested protein spots, identified by 4700 Proteomics Analyzer mass spectrometer analysis identified six lysine acetylation of protein, 8 lysine acetylation sites. The other is the use of immunoprecipitation enriched protein in whole liver protein acetylation, with the LTQ-Orbitrap mass spectrometry identification, identified 91 protein lysine acetylation, 216 lysine acetylation sites . To 97 in the identification of acetylated proteins, the majority have not been reported in the literature. We identified the biological functions of proteins, physiological processes, subcellular localization category. In addition, we use Q-Star flight mass spectrometer (ESI source) were mice liver threonine and serine protein acetylation on the identification of post-translational modification, identified 18 acetylated threonine protein post-translational modifications, 21 threonine acetylation site, 25 acetylated protein serine, serine 30 acetylation site, a great complement to the data. Chapter IV of this thesis acetylation posttranslational modifications done a summary of the work of this thesis were discussed. This study is the first to incorporate two types of biological mass spectrometry technology for small scale Shushu liver samples were acetylated histone modifications spectrum identification and analysis, for subsequent functional analysis of protein acetylation and its relationship with major diseases and cancer-related research provides a very good database reference.

Identification of protein acetylation

Abstract

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