病毒感染蛋白质组学研究进展

病毒的侵入会导致宿主细胞蛋白表达模式的改变,这种改变将影响宿主细胞的正常生理功能并决定病毒的致病进程和结果.因此,病毒感染蛋白质组学研究有助于揭示病毒与宿主的相互作用机制和病毒的分子致病机制,以及寻找病毒早期感染的分子标记、建立早期诊断方法、评价治疗效果和预后.本文介绍了病毒感染蛋白质组学研究技术、病毒诱导宿主细胞蛋白质组改变和病毒感染宿主血清差异蛋白质组等方面的研究进展.     

病毒蛋白质组学及其应用

病毒蛋白质组学是蛋白质组学研究技术在病毒学领域的应用,其研究方法主要是基于质谱鉴定的电泳分离或色谱分离技术。病毒蛋白质组学的研究可以补充基因组注释、纯化单一的病毒成分、研究病毒与其宿主细胞蛋白的相互作用、识别病毒作用的靶位点、鉴定病毒感染的致病因子及病毒的进化关系、识别病毒的免疫源性蛋白。病毒蛋白质组的研究有助于对病毒致病性的了解,加速新的诊断方法及治疗药物的研制,增强对病毒的生物防御。由于一些技术及主观因素的影响,病毒蛋白质组的研究是很有限的,这是一个亟待重视并增强的领域。     

种子蛋白质组的研究进展

蛋白质组学是通过对全套蛋白质动态的研究,来阐明生物体、组织、细胞和亚细胞全部蛋白质的表达模式及功能模式。大量可用的核苷酸序列信息和灵敏高速的质谱鉴定技术,使得蛋白质组学方法为分析模式植物和农作物的复杂功能开辟了新的途径。目前,种子蛋白质组研究主要集中在两个方面：一方面是鉴定尽可能多的蛋白,以创建种子特定生命时期的蛋白质组参照图谱;另一方面主要集中在差异蛋白质组,通过比较分析不同蛋白质组,以探明关键功能蛋白。该文综述了近年来种子蛋白质组的研究进展,内容包括种子发育过程中蛋白质组的变化,与种子休眠/萌发相关的蛋白质组、翻译后修饰蛋白质组、细胞与亚细胞差异蛋白质组以及环境因子对种子蛋白质组的影响;并对种子蛋白质组研究的热点问题进行了展望。     

种子蛋白质组的研究进展

蛋白质组学是通过对全套蛋白质动态的研究, 来阐明生物体、组织、细胞和亚细胞全部蛋白质的表达模式及功能模式。大量可用的核苷酸序列信息和灵敏高速的质谱鉴定技术, 使得蛋白质组学方法为分析模式植物和农作物的复杂功能开辟了新的途径。目前, 种子蛋白质组研究主要集中在两个方面: 一方面是鉴定尽可能多的蛋白, 以创建种子特定生命时期的蛋白质组参照图谱; 另一方面主要集中在差异蛋白质组, 通过比较分析不同蛋白质组, 以探明关键功能蛋白。该文综述了近年来种子蛋白质组的研究进展, 内容包括种子发育过程中蛋白质组的变化, 与种子休眠/萌发相关的蛋白质组、翻译后修饰蛋白质组、细胞与亚细胞差异蛋白质组以及环境因子对种子蛋白质组的影响; 并对种子蛋白质组研究的热点问题进行了展望。     

蛋白质组学技术在感染研究中的应用

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。     

蛋白质组学在干细胞研究中的应用

蛋白质组学技术通过整合多项技术来分析生物体的全部蛋白质成分,通过考察不同状态下细胞或组织蛋白质组的变化情况来了解细胞活动的分子机理。干细胞分化过程中受外界条件的影响其蛋白表达模式也表现出一定的差异,对干细胞分化过程中进行蛋白质组学研究将有利于从蛋白质分子水平上阐明干细胞的分化机理。本文对蛋白质组学及其在干细胞研究中的应用加以评述。     

基于质谱的蛋白质组学信号网络研究进展

细胞信号转导网络调控着所有细胞和器官的生物学过程。以往信号转导网络的研究主要采用一些生物化学方法开展,如抗体技术。目前,基于质谱的大规模蛋白质组学研究可以在翻译后修饰、蛋白质互作及蛋白质表达水平上,系统地研究信号转导事件。基于蛋白质组学的大规模信号转导的研究将改变我们对信号转导网络的理解。从蛋白质组翻译后修饰、蛋白质互作及蛋白质表达3个方面综述了质谱在信号转导方面的研究。     

iTRAQ技术在真菌研究中的应用进展

iTRAQ技术是一种新的、功能强大的、可以最多同时比较8种不同样品中蛋白质相对或绝对含量的蛋白质组学方法,结合多维液相色谱和串联质谱分析,iTRAQ技术已成为差异蛋白质组学定量研究的主要工具之一。而真菌的致病作用是多种蛋白质共同参与的真菌?宿主相互作用的复杂过程,因此整体、定量地分析真菌致病过程中的差异表达蛋白质谱,对于研究真菌的致病机制具有重要作用。该文重点就iTRAQ技术在真菌研究中的应用进展进行综述。     

细胞因子诱导外周血单个核细胞的转录和蛋白表达研究

采用干扰素-γ、抗CD3单克隆抗体和IL-2体外诱导扩增外周血单个核细胞成为cIK细胞,并于诱导培养前及培养第15d时分别收集细胞样本。在对培养前后细胞的增殖、形态及表面标志变化检测的同时。提取总蛋白进行定量、双向电泳和银染。利用ImageMasterTM软件对培养前后表达相同和不同的蛋白质点进行分析,并选择其中24个蛋白质点进行质谱鉴定。对于部分培养前后具有代表性的蛋白,进一步采用qPCR技术分析其的转录情况。结果表明,培养前后细胞的蛋白质组学特征是完全不同的,相同表达蛋白点主要与基因的转录因子和细胞骨架相关,诱导后特异表达蛋白主要与细胞生长、增殖相关。虽然在转录与蛋白水平上呈现出部分负相关现象,由于蛋白质组才是基因表达的最终形式,结合蛋白差异研究结果提示,经细胞因子诱导后,CIK细胞的大量扩增与细胞内蛋白表达改变相关。     

蛋白质组学技术在病毒学研究中的应用

蛋白质组学是门研究细胞和组织中蛋白质的存在及其结构、翻译后修饰、表达水平,最终阐述蛋白质群体功能的学科[1],它不仅是对机体产生的所有蛋白进行系统分离解析,更重要的是对这些蛋白质如何行使功能作出解释.利用先进的蛋白质组学技术研究病毒及相闰关宿主蛋白的表达、翻译后修饰、定位、相互作用和功能进行系统研究,有助于深入了解病素毒复复制、与宿主相互作用以及引发疾病的机制。     

Biomedical applications of protein chips

The development of microchips involving proteins has accelerated within the past few years. Although DNA chip technologies formed the precedent, many different strategies and technologies have been used because proteins are inherently a more complex type of molecule. This review covers the various biomedical applications of protein chips in diagnostics, drug screening and testing, disease monitoring, drug discovery (proteomics), and medical research. The proteomics and drug discovery section is further subdivided to cover drug discovery tools (on-chip separations, expression profiling, and antibody arrays), molecular interactions and signaling pathways, the identification of protein function, and the identification of novel therapeutic compounds. Although largely focused on protein chips, this review includes chips involving cells and tissues as a logical extension of the type of data that can be generated from these microchips.     

Proteomics technologies in endometriosis

Endometriosis is a common disorder that is associated with infertility and pelvic pain. Diagnosis is based on the visualization of endometriotic lesions during surgery as no reliable serum marker is currently available. The etiology of endometriosis is largely unknown. Over the last 20 years, several proteomics technologies have been used to research novel proteins with a potential etiological role in endometriosis, and to identify candidate serum markers for this condition. While some molecules identified by proteomics technologies may have a relevant role in the pathogenesis of endometriosis, the research of potential serum markers for this condition is still far from any clinical application. This review summarizes the state of the art and potential applications of proteomics in endometriosis research.     

翻译后修饰蛋白质组学研究的技术策略

 蛋白质组学早期研究的绝大部分工作是在关注细胞不同生长时期或是疾病、分裂素刺激下的蛋白质表达水平变化.然而,许多至关重要的生命进程不仅由蛋白质的相对丰度控制,更重要的是被那些时空特异分布的可逆翻译后修饰控制的,揭示翻译后修饰发生规律是理解蛋白质复杂多样的生物功能的一个重要前提.由于翻译后修饰蛋白质在样本中含量低且动态范围广,其相关研究极具挑战性,亲和富集、多维分离等技术与生物质谱的结合为翻译后修饰蛋白质组学的发展提供了契机,目前,已进行规模化研究的蛋白质翻译后修饰主要有四大类,其中磷酸化和糖基化研究较多.本文针对大规模翻译后的修饰蛋白质的分析策略和技术路线,如蛋白质的磷酸化修饰, 糖基化修饰, 泛素化修饰,基于蛋白质氧化还原状态进行的氧化还原修饰和其它修饰像乙酰化、甲基化、脂基化修饰等进行了综述.     

Clinical proteomics: present and future prospects

Advances in proteomics technology offer great promise in the understanding and treatment of the molecular basis of disease. The past decade of proteomics research, the study of dynamic protein expression, post-translational modifications, cellular and sub-cellular protein distribution, and protein-protein interactions, has culminated in the identification of many disease-related biomarkers and potential new drug targets. While proteomics remains the tool of choice for discovery research, new innovations in proteomic technology now offer the potential for proteomic profiling to become standard practice in the clinical laboratory. Indeed, protein profiles can serve as powerful diagnostic markers, and can predict treatment outcome in many diseases, in particular cancer. A number of technical obstacles remain before routine proteomic analysis can be achieved in the clinic; however the standardisation of methodologies and dissemination of proteomic data into publicly available databases is starting to overcome these hurdles. At present the most promising application for proteomics is in the screening of specific subsets of protein biomarkers for certain diseases, rather than large scale full protein profiling. Armed with these technologies the impending era of individualised patient-tailored therapy is imminent. This review summarises the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality.     

Proteomics and peptidomics in fundamental and applied medical studies

The review is focused on current issues of biomedical proteomics and peptidomics. The main attention is paid to modem proteomics technologies applied in medical research--extraction, detection and data analysis techniques. The use of chromatography, mass spectrometry and chromato mass spectrometry in proteogenomic, biomedical studies and biomarker discovery is discussed in detail.     

Proteomics and leukocytes: an approach to understanding potential molecular mechanisms of inflammatory responses

Leukocytes play an important role in the progression of disease and leukocyte-derived proteins are associated with the pathogenesis of the disease. Leukocyte activation causes production of inflammatory mediators, over-expression of cell surface adhesion molecules, and an increase in migration and infiltration, phagocytosis, and degranulation, as well as receptor phosphorylation and signal transduction. An increasing number of studies on the application of leukocyte proteomics have appeared in mapping protein profiles of inflammatory cells, contributing to the understanding of potential mechanisms involved in leukocyte function. Together with improvements in proteomic technology in leukocyte research, leukocyte proteomic analysis becomes more simple, rapid, flexible, sensitive, and specific. This enables proteomic investigation of activated or non-activated leukocytes to be highly focused on defined suborgans or specific signaling pathways. Research in leukocyte proteomics is progressing from fingerprinting to functioning, human cell lines to primary leukocytes, non-activated cells to inflammatory mediator-stimulated cells, in vitro culture to in vivo challenge, and animal models to human disease. A number of newly identified proteins from leukocyte proteomics may offer new mechanism-orientated targets for drug discovery and development.     

A streamlined approach to high-throughput proteomics

Proteomics has rapidly become an important tool for life science research, allowing the integrated analysis of global protein expression from a single experiment. To accommodate the complexity and dynamic nature of any proteome, researchers must use a combination of disparate protein biochemistry techniques, often a highly involved and time-consuming process. Whilst highly sophisticated, individual technologies for each step in studying a proteome are available, true high-throughput proteomics that provides a high degree of reproducibility and sensitivity has been difficult to achieve. The development of high-throughput proteomic platforms, encompassing all aspects of proteome analysis and integrated with genomics and bioinformatics technology, therefore represents a crucial step for the advancement of proteomics research. ProteomIQ? (Proteome Systems) is the first fully integrated, start-to-finish proteomics platform to enter the market. Sample preparation and tracking, centralized data acquisition and instrument control, and direct interfacing with genomics and bioinformatics databases are combined into a single suite of integrated hardware and software tools, facilitating high reproducibility and rapid turnaround times. This review will highlight some features of ProteomIQ, with particular emphasis on the analysis of proteins separated by 2D polyacrylamide gel electrophoresis.     

A streamlined approach to high-throughput proteomics

Proteomics has rapidly become an important tool for life science research, allowing the integrated analysis of global protein expression from a single experiment. To accommodate the complexity and dynamic nature of any proteome, researchers must use a combination of disparate protein biochemistry techniques, often a highly involved and time-consuming process. Whilst highly sophisticated, individual technologies for each step in studying a proteome are available, true high-throughput proteomics that provides a high degree of reproducibility and sensitivity has been difficult to achieve. The development of high-throughput proteomic platforms, encompassing all aspects of proteome analysis and integrated with genomics and bioinformatics technology, therefore represents a crucial step for the advancement of proteomics research. ProteomIQ (Proteome Systems) is the first fully integrated, start-to-finish proteomics platform to enter the market. Sample preparation and tracking, centralized data acquisition and instrument control, and direct interfacing with genomics and bioinformatics databases are combined into a single suite of integrated hardware and software tools, facilitating high reproducibility and rapid turnaround times. This review will highlight some features of ProteomIQ, with particular emphasis on the analysis of proteins separated by 2D polyacrylamide gel electrophoresis.     

Peptidomics, current status

Characterisation of the complement of expressed proteins from a single genome is a central focus of the evolving field of proteomics. Traditional proteomics technologies were developed in the 20th century and are based on two-dimensional electrophoresis or multidimensional liquid chromatography. These facilitated functional genomics analysis, but they currently represent a significant bottleneck to progress in this area. We are now witnessing the development of novel alternative technologies for use in expression proteomics research. This review aims to familiarise the reader with the principles underlying the peptidomics approaches to proteomics research and provide examples of their applications.     

自组装蛋白质芯片——功能蛋白质组学的新工具

传统的蛋白质芯片制备需要进行繁琐的蛋白质表达与纯化。同时,由于蛋白质活性不稳定,蛋白质芯片不宜长期保存。新一代自组装蛋白质芯片,利用无细胞表达体系和DNA固定技术,能够将蛋白质即时、原位表达并固定在芯片上,有效地解决了传统蛋白质芯片的制备和保存问题。目前自组装蛋白质芯片已初步用于大规模蛋白-蛋白质相互作用的筛选,以及鉴定免疫优势抗原等研究。该文介绍了近年自组装蛋白质芯片技术的进展和应用研究。