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介绍蛋白质组学研究的背景;阐述数据挖掘的原理、方法,重点讲述数据挖掘技术在蛋白质组学研究中取得的新的进展。最后,对数据挖掘目前存在的问题作分析,并对它的发展的前景作展望。 相似文献
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蛋白质组学是旨在研究蛋白质表达谱和蛋白质与蛋白质之间相互作用的新的领域。蛋白质组学的研究必须依赖高通量、自动化程度很高的技术。双向电泳、液相色谱和生物质谱技术的发展推动了蛋白质组学的研究。蛋白质组学为疾病发病机制的研究提供了新的思路和方法 ,本文重点介绍了蛋白质组学技术在心血管疾病研究中的应用 相似文献
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现代质谱技术在蛋白质组学中的应用及其最新进展 总被引:1,自引:0,他引:1
简述了蛋白质组学的概念、内容和意义,重点综述了现代质谱技术在蛋白质组学中的应用,主要包括蛋白质和肽段的鉴定和定量、蛋白质翻译后修饰的鉴定和蛋白质间相互作用的检测等。随着新的高质量精确度、分辨率、灵敏度和通量质谱仪的出现,现代质谱技术在蛋白质组学中的应用将越来越广泛,并给蛋白质组学研究带来新的机遇。 相似文献
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蛋白质组学的目标在于阐明特定生物体、组织、细胞或亚细胞结构中全部蛋白质的表达模式和功能模式,其技术平台由高通量的蛋白质分离技术、鉴定技术和生物信息学组成。在许多研究领域,蛋白质组学技术为阐明疾病过程和生命现象的分子机制提供了全面、网络和动态的蛋白质组信息。感染是重要的基本致病因素之一,蛋白质组学的研究策略和技术方法有利于快速分离鉴定病原体蛋白质组、宿主免疫细胞蛋白质组、感染相关蛋白、疫苗候:选抗原蛋白、生物标志物和药物靶标,从而明显加快病原体、宿主反应、感染发病机制以及感染预防、诊断和治疗等相关研究的进程。 相似文献
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微循环遍及全身,直接参与机体物质、能量与信息传递,与心、脑及外周血管疾病、糖尿病、结缔组织病及创伤、感染、休克等病理过程的发生、发展、疗效及预后判断关系密切,其功能改变涉及多种细胞因子与蛋白质组分之间动态、复杂、精细的相互作用。蛋白质组学作为一门研究细胞蛋白质的组成及其活动规律的新兴学科,有助于阐明生理或病理条件下的微循环改变的分子机制。本文综述近年来蛋白质组学技术进展及其在微循环研究中的应用。 相似文献
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蛋白质组学技术在神经系统疾病研究中的作用 总被引:1,自引:0,他引:1
双向凝胶电泳和质谱等方法都是蛋白质组学(Proteomics)技术的重要方法。应用蛋白质组学技术可以同时研究大量蛋白质的功能、组成,多样性及其动态变化。神经科学的许多问题可以借助于这个新的工具平台获得解决,因此,蛋白质组学的发展,将为神经疾病发病机制的深入研究,以及相关的药物开发提供一个崭新的发展机遇。 相似文献
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《Expert review of proteomics》2013,10(4):401-409
2D polyacrylamide gel electrophoresis has been the traditional workhorse of proteomics, allowing for the resolution of several thousand proteins in a single gel. Difference gel electrophoresis is an emerging technology that allows for accurate quantification with statistical confidence while controlling for nonbiologic variation, and also increases the dynamic range and sensitivity of traditional 2D polyacrylamide gel electrophoresis. With inclusion of an internal standard formed from equal amounts of every sample in an experiment, difference gel electrophoresis technology also allows for repetitive measurements and multivariable analyses to be quantitatively analyzed in one co-ordinated experiment, yielding statistically-significant changes in protein expression related to many disease states. This technique promises to be an important tool in clinical proteomics and the study of the mechanism of disease, investigating diagnostic biomarkers and pinpointing novel therapeutic targets. 相似文献
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Lincet H Guével B Pineau C Allouche S Lemoisson E Poulain L Gauduchon P 《Journal of Proteomics》2012,75(4):1157-1169
Ovarian cancer is the fifth most frequent cause of cancer death in women. Emergence of chemoresistance in the course of treatments with platinum drugs is in part responsible for therapeutic failures. In order to improve the understanding of the complex mechanisms involved in acquired platinum chemoresistance, we decided to compare the basal protein expression profile of the platinum-sensitive cell line OAW42 and that of its resistant counterpart OAW42-R by a proteomic approach. Reversed-phase HPLC pre-fractionated extracts from both cell lines were subjected to 2D-DIGE coupled to mass spectrometry (MS). Forty eight differentially expressed proteins were identified, 39 being up-regulated and 19 down-regulated in OAW42-R versus OAW42 cells. From the current knowledge on biological activities of most differentially expressed proteins, it can be inferred that the acquisition of resistance was associated with a global reorganization of biochemical pathways favoring the production of precursors for biosynthesis, and with the mobilization of macromolecule quality control mechanisms, preserving RNA and protein integrity under damage-inducing conditions. 相似文献
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Kondo T 《BMB reports》2008,41(9):626-634
Novel cancer biomarkers are required to achieve early diagnosis and optimized therapy for individual patients. Cancer is a disease of the genome, and tumor tissues are a rich source of cancer biomarkers as they contain the functional translation of the genome, namely the proteome. Investigation of the tumor tissue proteome allows the identification of proteomic signatures corresponding to clinico-pathological parameters, and individual proteins in such signatures will be good biomarker candidates. Tumor tissues are also a rich source for plasma biomarkers, because proteins released from tumor tissues may be more cancer specific than those from non-tumor cells. Two-dimensional difference gel electrophoresis (2D-DIGE) with novel ultra high sensitive fluorescent dyes (CyDye DIGE Fluor satulation dye) enables the efficient protein expression profiling of laser-microdissected tissue samples. The combined use of laser microdissection allows accurate proteomic profiling of specific cells in tumor tissues. To develop clinical applications using the identified biomarkers, collaboration between research scientists, clinicians and diagnostic companies is essential, particularly in the early phases of the biomarker development projects. The proteomics modalities currently available have the potential to lead to the development of clinical applications, and channeling the wealth of produced information towards concrete and specific clinical purposes is urgent. 相似文献
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Tokutomi Y Araki N Kataoka K Yamamoto E Kim-Mitsuyama S 《Biochemical and biophysical research communications》2007,364(4):822-830
To study early changes in angiotensin II (Ang II)-induced signaling with post-translational modifications, we analyzed proteins from cultured human coronary smooth muscle cells stimulated with Ang II, using two-dimensional difference gel electrophoresis (2D-DIGE) combined with ProQ Diamond and SYPRO Ruby staining, followed by mass spectrometry or Western blotting. Among 40 proteins identified, peroxiredoxin 2 (Prx2) was oxidized and 58 kDa glucose-regulated protein (GRP58) was phosphorylated after 5 min of Ang II (1 μM) stimulation. Valsartan, a selective Ang II type 1 (AT1) receptor blocker, and N-acetylcysteine, an antioxidant, inhibited both of these modifications, indicating the contribution of AT1 receptor and reactive oxygen species to oxidation of Prx2 and phosphorylation of GRP58 by Ang II. 相似文献
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Francisco-Jose Fernandez-Gomez Fanny Jumeau Maxime Derisbourg Sylvie Burnouf Hélène Tran Sabiha Eddarkaoui Hélène Obriot Virginie Dutoit-Lefevre Vincent Deramecourt Valérie Mitchell Didier Lefranc Malika Hamdane David Blum Luc Buée Valérie Buée-Scherrer Nicolas Sergeant 《Journal of visualized experiments : JoVE》2014,(86)
Two-dimensional gel electrophoresis (2DE) is a powerful tool to uncover proteome modifications potentially related to different physiological or pathological conditions. Basically, this technique is based on the separation of proteins according to their isoelectric point in a first step, and secondly according to their molecular weights by SDS polyacrylamide gel electrophoresis (SDS-PAGE). In this report an optimized sample preparation protocol for little amount of human post-mortem and mouse brain tissue is described. This method enables to perform both two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mini 2DE immunoblotting. The combination of these approaches allows one to not only find new proteins and/or protein modifications in their expression thanks to its compatibility with mass spectrometry detection, but also a new insight into markers validation. Thus, mini-2DE coupled to western blotting permits to identify and validate post-translational modifications, proteins catabolism and provides a qualitative comparison among different conditions and/or treatments. Herein, we provide a method to study components of protein aggregates found in AD and Lewy body dementia such as the amyloid-beta peptide and the alpha-synuclein. Our method can thus be adapted for the analysis of the proteome and insoluble proteins extract from human brain tissue and mice models too. In parallel, it may provide useful information for the study of molecular and cellular pathways involved in neurodegenerative diseases as well as potential novel biomarkers and therapeutic targets. 相似文献
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In order to explore the molecular mechanism underlying self-incompatibility (SI) in the apricot (Prunus armeniaca L.) at the proteome level, we examined the style proteomes at different stages of flower development: small bud, big bud, 24h after self-pollination and 24h after cross-pollination with cultivar Badanshui in the SI apricot cultivar Xinshiji and the self-compatible (SC) apricot cultivar Katy by 2D fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS). About 1500 style protein spots were detected; 66 were expressed differently in the four stages in Xinshiji. About 1600 style protein spots were detected; 143 were expressed differently in the four stages of flower development in Katy. In Xinshiji, one protein was expressed specifically, four proteins showed up-regulated expression and twenty-nine proteins showed down-regulated expression in the cross-pollinated style compared to the self-pollinated style. Thirteen proteins were identified unambiguously. In Katy, three proteins were expressed specifically, five proteins showed up-regulated expression and thirteen proteins showed down-regulated expression in the cross-pollinated style compared to self-pollinated style. Seven proteins were identified unambiguously. The different reactions of the style at the proteomic level were triggered in Xinshiji and Katy by self pollen and non-self pollen. 相似文献
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Lab-on-a-chip: applications in proteomics 总被引:17,自引:0,他引:17
Mouradian S 《Current opinion in chemical biology》2002,6(1):51-56
Recent advances in chip-based separation of proteins provide methods that are faster and more convenient than conventional gel electrophoresis. Rapid and automated protein sizing on a chip is at the commercial stage and first attempts have been made to perform two-dimensional separation on a chip. Numerous designs have been described to interface a microfluidic chip to a mass spectrometer. Impressive integration efforts are demonstrated by the ability to perform on-chip trypsin digestion, separation and injection into a mass spectrometer with a single device. 相似文献
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Kikuta K Kubota D Saito T Orita H Yoshida A Tsuda H Suehara Y Katai H Shimada Y Toyama Y Sato K Yao T Kaneko K Beppu Y Murakami Y Kawai A Kondo T 《Journal of Proteomics》2012,75(4):1089-1098
Gastrointestinal stromal tumor (GIST) is the most common mesenchymal malignancy of the gastrointestinal tract, comprising a wide spectrum from a curable disorder to highly malignant disease. GIST is characterized by tyrosine kinase mutations, and molecular targeting therapies against these abnormal enzymes require prognostic biomarkers. To identify candidate prognostic biomarkers, we examined proteomic features corresponding to metastasis after surgery. Using two-dimensional difference gel electrophoresis with a large format gel, we compared the primary tumor tissues of GIST patients free of metastasis for two years after surgery (eight cases) with those of patients who developed metastasis within one year after surgery (nine cases). We found the intensities of 38 protein spots to differ significantly between the two groups. Mass spectrometric protein identification revealed that these corresponded to 25 unique genes. Immunohistochemical validation demonstrated ATP-dependent RNA helicase DDX39 to be significantly associated with metastasis and poor clinical outcomes in a group of 72 GIST patients. In conclusion, we have established a novel prognostic utility of ATP-dependent RNA helicase DDX39 in GIST.ATP-dependent RNA helicase DDX39, a novel biomarker for GIST likely to be associated with metastatic disease, can identify patients likely to benefit from new therapeutic strategies such as tyrosine kinase inhibitors. 相似文献
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Kakhniashvili DG Griko NB Bulla LA Goodman SR 《Experimental biology and medicine (Maywood, N.J.)》2005,230(11):787-792
Quantitative changes in the red blood cell membrane proteome in sickle cell disease were analyzed using the two-dimensional fluorescence difference gel electrophoresis 2D-DIGE technique. From over 500 analyzed two-dimensional gel spots, we found 49 protein gel spots whose content in sickle cell membranes were changed by at least 2.5-fold as compared to control cells. In 38 cases we observed an increase and in 11 cases a decrease in content in the sickle cell membranes. The proteins of interest were identified by in-gel tryptic digestion followed by liquid chromatography in line with tandem mass spectrometry. From 38 analyzed gel spots, we identified 44 protein forms representing different modifications of 22 original protein sequences. The majority of the identified proteins fall into small groups of related proteins of the following five categories: actin accessory proteins--four proteins, components of lipid rafts--two proteins, scavengers of oxygen radicals--two proteins, protein repair participants--six proteins, and protein turnover components--three proteins. The number of proteins whose content in sickle RBC membrane is decreased is noticeably smaller, and most are either components of lipid rafts or actin accessory proteins. Elevated content of protein repair participants as well as oxygen radical scavengers may reflect the increased oxidative stress observed in sickle cells. 相似文献