蛋白质组学及其技术进展

蛋白质组学是系统生物学重要基础和组成部分之一，近年来其研究技术进展迅猛。本文分析了蛋白质组学研究的必要性、长期性、复杂性和挑战性，重点综述了蛋白质组学一些研究技术及其在近年的进展。     

蛋白质组学 科研进行时

行业视角：人类基因组计划取得成功后，生命科学研究进入后基因组时代，而其中与生命状态息息相关的蛋白质表达和蛋白质结构分析及蛋白质作用位点研究已成为热点和前沿，如药物分子靶向性治疗、药物前导化合物筛选、疾病相关细胞生物学研究、新生儿遗传疾病和易发易感疾病的相关研究，干细胞分化研究等等，而发现新的生物标志物（Biomarker）是目前生命科学与蛋白质组研究的重要目标与任务之一。[第一段]     

蛋白质组学中的有机质谱

本文综述了基体辅助激光解析电离飞行时间质谱,电喷雾电离串联质谱的发展及其在蛋白质组学中的应用。     

基于质谱的蛋白质组学方法新进展

蛋白质组学是后基因组时代的科研热点,其研究方向主要涵盖蛋白质表达谱、蛋白质翻译后修饰谱、蛋白质相互作用谱和单细胞蛋白质组等定性分析以及相对和绝对定量分析,现已广泛应用于生物、医药及病理研究中.由于质谱具有灵敏、准确、高通量等特点,是蛋白质组学研究必不可少的工具.本文综述了近年来基于质谱技术的蛋白质组鉴定及定量方法,并展...     

蛋白质组学面临的新问题

蛋白质组学领域仅仅经历了十年的发展历程.虽然它还有许多棘手的问题尚待解决,但是研究人员坚信蛋白质组学能够发现新的药靶和生物标记物,从而可以改进诊断和治疗过程.本文选择此领域内几家瑞士公司的工作进行评述.     

蛋白质组学面临的新问题

蛋白质组学领域仅仅经历了十年的发展历程。虽然它还有许多棘手的问题尚待解决，但是研究人员坚信蛋白质组学能够发现新的药靶和生物标记物，从而可以改进诊断和治疗过程。本文选择此领域内几家瑞士公司的工作进行评述。     

植物逆境生物学中的蛋白质组学技术运用

逆境胁迫会对植物生长发育造成制约,并且对作物产量、质量等造成影响,植物应答胁迫分子机理的分析为人们尤为重视的内容。植物蛋白质组学的研究能够系统性的揭露在胁迫条件不同时,植物蛋白质表达的情况,以此对基于环境胁迫下植物基因表达调控机制进行掌握。以此,简单分析了蛋白质组学技术在植物逆境生物学中的使用。     

SELDI蛋白质芯片技术在蛋白质组学中的应用

本文介绍了SELDI(Suface-enhanced Laser Desorption/Ionization)蛋白质芯片技术的工作原理,讨论了不同类型的蛋白质芯片在蛋白质组学中的应用。     

蛋白质组学研究中的多维液相分离系统

在后基因组时代,蛋白质组学成为新的研究热点。多维液相分离系统具有分离样品动态范围宽、分辨率高、峰容量大、分析速度快、自动化程度高等优点,因而近年来在蛋白质组学的研究中受到了越来越多的关注。本文根据目前已发表的文章,对二维液相色谱(2D-LC)、二维液相色谱-毛细管电泳(2D-LC-CE)、二维毛细管电泳(2D-CE)以及它们在蛋白质组学中的应用进行了系统的综述。     

创新的蛋白质组学质谱应用新技术——SWATH技术

AB SCIEX公司在2011年美国第59届质谱年会上新推出了SWATH（Sequential Windowed Acquisition of all Theoretical fragment ions,SWATH）采集技术,该技术是与苏黎世联邦理工学院Ruedi Aebersold博士及其团队共同开发的。SWATH采集模式为研究蛋白质组学的用户带来了创新的技术,科学家利用该技术可以在一次进样分析中即可获得复杂蛋白质组样品中每个肽段的数据。SWATH采集模式是AB SCIEX的MS/MSALL技术的延伸,此功能只能在TripleTOFTM5600系统上完成,作为对MRMAtlas的完美补充,为未来蛋白质组学研究提供了一种全新的工作流程,预计将产生令人震撼的蛋白质组学新发现。     

Cancer proteomics

Cancer presents high mortality and morbidity globally, largely due to its complex and heterogenous nature, and lack of biomarkers for early diagnosis. A proteomics study of cancer aims to identify and characterize functional proteins that drive the transformation of malignancy, and to discover biomarkers to detect early‐stage cancer, predict prognosis, determine therapy efficacy, identify novel drug targets, and ultimately develop personalized medicine. The various sources of human samples such as cell lines, tissues, and plasma/serum are probed by a plethora of proteomics tools to discover novel biomarkers and elucidate mechanisms of tumorigenesis. Innovative proteomics technologies and strategies have been designed for protein identification, quantitation, fractionation, and enrichment to delve deeper into the oncoproteome. In addition, there is the need for high‐throughput methods for biomarker validation, and integration of the various platforms of oncoproteome data to fully comprehend cancer biology. © 2012 Wiley Periodicals, Inc. Mass Spec Rev 31:583–605, 2012     

光声成像及其在生物研究中的应用

作为新型的活体检测设备,商业化的光声成像装置走向市场。本文综述光声成像装置的原理、基本构成、系统集成及发展趋势,尤其关注其在生物领域的应用。     

Microwave-assisted proteomics

State-of-the-art proteomic analysis has recently undergone a rapid evolution; with more high-throughput analytical instrumentation and informatic tools available, sample preparation is becoming one of the rate-limiting steps in protein characterization workflows. Recently several protocols have appeared in the literature that employ microwave irradiation as a tool for the preparation of biological samples for subsequent mass spectrometric characterization. Techniques for microwave-assisted bio-catalyzed reactions (including sample reduction and alkylation, enzymatic and chemical digestion, removal and analysis of post-translational modifications and characterization of enzymes and protein-interaction sites) are described. This review summarizes the various approaches undertaken, instrumentation employed, and reduction in overall experimental time observed when microwave assistance is applied.     

Urinary exosomes and proteomics

A number of highly abundant proteins in urine have been identified through proteomics approaches, and some have been considered as disease-biomarker candidates. These molecules might be clinically useful in diagnosis of various diseases. However, none has proven to be specifically indicative of perturbations of cellular processes in cells associated with urogenital diseases. Exosomes could be released into urine which flows through the kidney, ureter, bladder and urethra, with a process of filtration and reabsorption. Urinary exosomes have been recently suggested as alternative materials that offer new opportunities to identify useful biomarkers, because these exosomes secreted from epithelial cells lining the urinary track might reflect the cellular processes associated with the pathogenesis of diseases in their donor cells. Proteomic analysis of such urinary exosomes assists the search of urinary biomarkers reflecting pathogenesis of various diseases and also helps understanding the function of urinary exosomes in urinary systems. Thus, it has been recently suggested that urinary exosomes are one of the most valuable targets for biomarker development and to understand pathophysiology of relevant diseases.     

Lipolytic proteomics

Activity‐based proteomics (ABP) employs small molecular probes to specifically label sets of enzymes based on their shared catalytic mechanism. Given that the vast majority of lipases belong to the family of serine hydrolases and share a nucleophilic active‐site serine as part of a catalytic triad, activity‐based probes are ideal tools to study lipases and lipolysis. Moreover, the ability of ABP to highlight or isolate specific subproteomes results in a massive decrease of sample complexity. Thereby, in‐depth analysis of enzymes of interest with mass spectrometry becomes feasible. In this review, we cover probe design, technological developments, and applications of ABP of lipases, as well as give an overview of relevant identified proteins. © 2012 Wiley Periodicals, Inc. Mass Spec Rev 31:570–582, 2012     

分子影像技术在转化医学中的应用

转化医学是当前医学领域的前沿和热点,旨在建立基础研究与临床应用之间的直接通路,建立实验室研究与临床医学之间的桥梁,有着广阔的发展前景。分子影像技术能够通过特殊的成像手段与条件,反映机体组织、细胞及分子水平的代谢和功能状态的变化,不仅是疾病早期诊断的利器,在新药研发方面也有着巨大的应用前景,还可为个体化治疗提供一个很好的指导工具。因此,分子影像技术将是转化医学得以顺利实现的一个重要手段,其在转化医学中的应用将会带来新的机遇,同时也会有更多的挑战。     

Subcellular proteomics

The step from the analysis of the genome to the analysis of the proteome is not just a matter of numerical complexity in terms of variants of gene products that can arise from a single gene. A significant further level of complexity is introduced by the supramolecular organization of gene products because of protein-protein interactions or targeting of proteins to specific subcellular structures. There is currently no single proteome analysis strategy that can sufficiently address all levels of the organization of the proteome. To approach an appropriate analytical complement for the interrogation of the proteome at all of the levels at which it is organized, there emerges the need for a whole arsenal of proteomics strategies. The proteome analysis at the level of subcellular structures (that can be enriched by subcellular fractionation) represents an analytical strategy that combines classic biochemical fractionation methods and tools for the comprehensive identification of proteins. Among the key potentials of this strategy is the capability to screen not only for previously unknown gene products but also to assign them, along with other known, but poorly characterized gene products, to particular subcellular structures. Furthermore, the analysis at the subcellular level is a prerequisite for the detection of important regulatory events such as protein translocation in comparative studies. This review is meant to give an overview on recent key studies in the field of proteome analysis at the level of subcellular structures, and to highlight potentials and requirements.     

Phase contrast in Simultaneous Topography and Recognition imaging

The operation of a force microscope in Simultaneous Topography and Recognition (TREC) imaging mode is analyzed by means of numerical simulations. Both topography and recognition signals are analyzed by using a worm-like chain force as the specific interaction between the functionalized tip probe and the sample. The special feedback mechanism in this mode is shown to couple the phase signal to the presence of molecular recognition interactions even in absence of dissipation.