蛋白质组学在食管癌发生中的研究进展

随着人类基因组全序列草图的完成,从基因水平向蛋白质水平的深化,已成为生命科学研究的迫切需要和新的任务。蛋白质组学是研究蛋白质组成和动态变化的一门新兴学科,蛋白质组学研究技术已经应用于肿瘤的发生机制、诊断、治疗之中。从蛋白质整体水平上研究食管癌的发生与转移,寻找食管癌发生及转移相关的新的蛋白质、特异性的标志物及药物治疗的靶标,对食管癌的诊治将起到重要作用。现综述蛋白质组学研究在食管癌发生中的研究进展。     

蛋白质组学及其在结核病研究中的应用

蛋白质组学是整体、动态、定性、定量地研究蛋白质组成、功能和相互关系的一门新兴学科。蛋白质组学的核心技术是双向电泳和生物质谱。目前蛋白质组学已用于结核分枝杆菌蛋白质组成、T细胞抗原、耐药机制等方面的研究,旨在发现结核病早期诊断标志物,开发预防结核病的新疫苗,寻找新的抗结核药物。     

蛋白质组学技术与针灸防治胃溃疡效应

蛋白质组学是一门在蛋白质水平认识生命机制的学科.近年来,应用蛋白质组学技术开展针灸防治胃溃疡效应的研究日益受到重视.本文综述了目前针灸防治胃溃疡的蛋白质组学研究的相关进展,并就研究中存在的相关问题进行分析,认为利用蛋白质组技术研究针灸机制和调节效应是未来针灸研究的重要突破口和发展方向.     

弥漫性间质性肺疾病蛋白组学研究

蛋白质组学是指对蛋白质组整体水平进行研究.与传统研究单一蛋白质变化的方法 发相比,能获得整体蛋白质水平表达差异.弥漫性间质性肺疾病发病机制复杂,涉及广泛的细胞因子网络,以往大量研究集中于单个细胞因子水平,蛋白质组学技术以其高新系统量及整体水平比较差异优势成为新的研究方向.本文就蛋白质组学技术在肺间质疾病的研究应用进行综述.     

动脉粥样硬化不稳定斑块的蛋白质组学研究的进展

在动脉粥样硬化的研究中,蛋白质组学技术可直接研究蛋白质表达,在临床上广泛应用于生物标志物鉴定和药物靶点识别。本综述详细描述了蛋白质组学在鉴别不稳定斑块生物标志物中的研究进展,从斑块内出血的蛋白质组学、血小板的蛋白质组学、细胞外基质的蛋白质组学等方面进行了报道。     

肿瘤蛋白质组学与消化系肿瘤生物标记

肿瘤蛋白质组学是通过蛋白质组技术研究肿瘤细胞中蛋白质及其相互作用的新的学科。将蛋白质组学技术用于研究肿瘤的发病机制、寻找新的肿瘤诊断生物标记以及采用组织蛋白质组图谱进行早期诊断等早已引起了人们的广泛关注。肿瘤蛋白质组学可能会引起肿瘤临床实践的重大革命。本文对近年发现的肿瘤生物标记进行了回顾,分析了肿瘤蛋白质组学在肿瘤生物标记发展中的前景。     

蛋白质组学临床转化研究主要进展及在心脏研究领域应用的挑战和展望

进入新世纪以来,作为后基因组时代的蛋白质组学研究成为了前沿热点研究领域之一。蛋白质组学在中国的发展一直保持着与国际齐头并进的态势,甚至在部分领域引领蛋白质组学的发展。在国外,蛋白质组学临床转化研究主要进展集中于美国的临床蛋白质组肿瘤分析协会(CPTAC)发表的论文。本文综述了“中国人类蛋白质组计划(CHPP)”和CPTAC目前主要的研究成果,以及少量在心脏领域应用蛋白质组学的研究进展。无论是CHPP,还是CPTAC,目前主要关注的是肿瘤领域,蛋白质组学在心脏领域的研究依然处于起步阶段,本文提出了蛋白质组学技术在心脏研究领域应用的挑战和相应的解决方法,就目前蛋白质组学技术在心脏研究领域应用的现状加以综述和展望。     

蛋白质组学研究方法

随着人类功能基因组计划的实施和推进，以蛋白质组(Proteome)和蛋白质组学(Proteomics)的提出为标志，人类从蛋白质水平全面揭示生命本质的研究进入一个全新的领域。“工欲善其事，必先利其器”，近年来蛋白质组学的迅猛发展与研究方法的改进和研究手段的提高是密不可分的。蛋白质组学的发展是伴随着蛋白质研究技术，尤其是双向凝胶电泳(2-D凝胶电泳)和新型质谱技术以及生物信息学的发展而发展的。本文将对主要蛋白质组学技术作一概述。     

蛋白质组学与胃肠相关心因性疾病

蛋白质组学以细胞、组织或机体内全部蛋白质为研究对象，研究其结构、功能和相互作用，具有高敏感性和特异性。蛋白质组学可发现精神疾病的特异性标记物，提高对精神疾病病理和发病机制的认识。采用蛋白质组学检测精神疾病的特异性蛋白质图谱，继而建立实验诊断模型，可为精神疾病的病因、诊断、分型、潜在药物靶点等的研究提供科学依据。本文就蛋白质组学与胃肠相关心因性疾病的关系作一综述。     

蛋白质芯片激光解析电离技术在肺癌研究中的应用

蛋白质芯片表面加强激光解析电离-飞行时间-质谱(SELDI-TOF-MS)技术是蛋白质组学研究的全新技术平台,进一步提高了蛋白质分离和鉴定的速度。并且在肺肿瘤生物标志物筛选、鉴别肺癌抗原和蛋白质指纹图谱方面取得突破性进展。今后该技术在肿瘤蛋白质组学研究中有更加广阔的应用前景。     

A tagging-via-substrate technology for detection and proteomics of farnesylated proteins

A recently developed proteomics strategy, designated tagging-via-substrate (TAS) approach, is described for the detection and proteomic analysis of farnesylated proteins. TAS technology involves metabolic incorporation of a synthetic azido-farnesyl analog and chemoselective derivatization of azido-farnesyl-modified proteins by an elegant version of Staudinger reaction, pioneered by the Bertozzi group, using a biotinylated phosphine capture reagent. The resulting protein conjugates can be specifically detected and/or affinity-purified by streptavidin-linked horseradish peroxidase or agarose beads, respectively. Thus, the technology enables global profiling of farnesylated proteins by enriching farnesylated proteins and reducing the complexity of farnesylation subproteome. Azido-farnesylated proteins maintain the properties of protein farnesylation, including promoting membrane association, Ras-dependent mitogen-activated protein kinase kinase activation, and inhibition of lovastatin-induced apoptosis. A proteomic analysis of farnesylated proteins by TAS technology revealed 18 farnesylated proteins, including those with potentially novel farnesylation motifs, suggesting that future use of this method is likely to yield novel insight into protein farnesylation. TAS technology can be extended to other posttranslational modifications, such as geranylgeranylation and myristoylation, thus providing powerful tools for detection, quantification, and proteomic analysis of posttranslationally modified proteins.     

酒精性肝病患者血浆外泌体差异蛋白的分析

目的筛选酒精性肝病(alcoholic liver disease, ALD)患者血浆外泌体中的差异蛋白,分析其功能及生物学过程,为ALD患者的治疗和诊断提供参考依据。方法选取2021年5月至2021年10月在首都医科大学附属北京佑安医院住院并确诊为ALD的患者和健康体检者各3例,超速离心分离提纯外泌体,提取蛋白,串联质谱标签(tandem mass tag, TMT)标记后,采用定量蛋白组学技术对两者血浆中的外泌体蛋白进行鉴定和定量分析,筛选出差异蛋白,并用生物信息学分析差异蛋白的功能及其参与的生物学过程。结果共鉴定到可定量蛋白质387种,以倍数上调>1.2倍或下调>1.2倍且P<0.05为标准筛选出差异蛋白27种,与健康对照组相比,ALD组上调蛋白15种、下调蛋白12种,生物信息学分析结果显示,这些蛋白主要参与了脂质的代谢、免疫反应和细胞的死亡等生物学过程,并与炎症反应、细胞的损伤和补体级联反应等信号通路密切相关。结论 TMT标记定量蛋白质组学技术筛选出的差异蛋白可能作为ALD早期诊断的血清学标志物及治疗靶点。     

Exosome Display technology: applications to the development of new diagnostics and therapeutics

Exosome Display is a novel methodology enabling the manipulation of exosome protein content. This technology stems from the identification of addressing domains that mediate the specific distribution of proteins on exosomes. More particularly, Lactadherin expressed in non-mammary gland tissue has been found to localize to exosomes via binding of its C1C2 domain to exosome lipids. Exosome Display of soluble antigens and extracellular domains of membrane proteins that are not naturally found on exosomes occurs upon fusion of proteins with the Lactadherin C1C2 domain. Exosome Display of native full-length membrane proteins can also be achieved by non-restricted expression or sampling of membrane proteins on exosomes. These novel findings enable us to manipulate exosome composition and tailor exosomes with new desirable properties. The Exosome Display technology is very versatile since soluble, membrane-bound, trans-membrane or multimeric antigens that are not naturally found on exosomes can now be efficiently expressed at their surface in a native conformation. The technology was applied to the generation of antibodies against tumor biomarkers such as HLA/peptide complex. This antibody method called ExoMAb can be used to generate antibodies against any drug target candidates, notably including G-protein coupled receptors. The potential of Exosome Display technology for developing a broad range of novel diagnostics and therapeutics is discussed.     

Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT)

In both normal and pathological states, cells respond rapidly to environmental cues by synthesizing new proteins. The selective identification of a newly synthesized proteome has been hindered by the basic fact that all proteins, new and old, share the same pool of amino acids and thus are chemically indistinguishable. We describe here a technology, based on the cotranslational introduction of azide groups into proteins and the chemoselective tagging of azide-labeled proteins with an alkyne affinity tag, to separate and identify, specifically, the newly synthesized proteins in mammalian cells. Incorporation of the azide-bearing amino acid azidohomoalanine is unbiased, not toxic, and does not increase protein degradation. As a first demonstration of the method, we report the selective purification and identification of 195 metabolically labeled proteins with multidimensional liquid chromatography in-line with tandem MS. Furthermore, in combination with leucine-based mass tagging, candidates were immediately validated as newly synthesized proteins. The identified proteins, synthesized in a 2-h window, possess a broad range of biochemical properties and span most functional gene ontology categories. This technology makes it possible to address the temporal and spatial characteristics of newly synthesized proteomes in any cell type.     

Comparative proteome analysis of peripheral blood mononuclear cells in systemic lupus erythematosus with iTRAQ quantitative proteomics

To identify and quantify protein profiles from peripheral blood mononuclear cells (PBMC) of systemic lupus erythematosus (SLE) patients with isobaric Tagging for Relative and Absolute protein Quantification (iTRAQ)–based proteomic technology and to find differentially expressed proteins in SLE. PBMC were collected from patients of six stable SLE, six active SLE, six rheumatoid arthritis (RA), and six healthy donors. After protein extraction and concentration, the pooled protein content was labeled with iTRAQ reagents and then subjected to multiple chromatographic fractionation and tandem mass spectrometry. ProteinPilot™ 3.0 software and a database of IPI (International Protein Index) human 3.62 were used for database searching and statistical analysis. A total of 452 proteins were identified. Of these, 67 unique proteins were observed twofold or more alteration in levels across groups. The proteins determined support existing knowledge and uncover novel biomarker candidates. These results indicate that iTRAQ-based technology can serve as a useful aid for identification and quantification proteins from PBMC.     

马尔尼菲篮状菌分泌蛋白的鉴定及功能预测分析

目的 鉴定马尔尼菲篮状菌(TM)的分泌蛋白,并初步分析其生物学功能.方法 基于质谱技术对TM菌株(ATCC18224)培养上清进行分泌蛋白组鉴定,利用生物信息学手段预测所鉴定蛋白的亚细胞定位和信号肽剪切位点,以及分析其生物学功能和信号通路.结果 通过2次独立重复实验,共鉴定得到21个分泌蛋白;功能注释结果显示分泌蛋白主...     

Pancreatic cancer proteome: the proteins that underlie invasion, metastasis, and immunologic escape

BACKGROUND & AIMS: Pancreatic cancer is a highly lethal disease that has seen little headway in diagnosis and treatment for the past few decades. The effective treatment of pancreatic cancer is critically relying on the diagnosis of the disease at an early stage, which still remains challenging. New experimental approaches, such as quantitative proteomics, have shown great potential for the study of cancer and have opened new opportunities to investigate crucial events underlying pancreatic tumorigenesis and to exploit this knowledge for early detection and better intervention. METHODS: To systematically study protein expression in pancreatic cancer, we used isotope-coded affinity tag technology and tandem mass spectrometry to perform quantitative proteomic profiling of pancreatic cancer tissues and normal pancreas. RESULTS: A total of 656 proteins were identified and quantified in 2 pancreatic cancer samples, of which 151 were differentially expressed in cancer by at least 2-fold. This study revealed numerous proteins that are newly discovered to be associated with pancreatic cancer, providing candidates for future early diagnosis biomarkers and targets for therapy. Several differentially expressed proteins were further validated by tissue microarray immunohistochemistry. Many of the differentially expressed proteins identified are involved in protein-driven interactions between the ductal epithelium and the extracellular matrix that orchestrate tumor growth, migration, angiogenesis, invasion, metastasis, and immunologic escape. CONCLUSIONS: Our study is the first application of isotope-coded affinity tag technology for proteomic analysis of human cancer tissue and has shown the value of this technology in identifying differentially expressed proteins in cancer.     

Ultrasound-assisted insulin delivery and noninvasive glucose sensing

Peptides and proteins are emerging as an increasingly important class of drugs as they become more readily available through improvement in recombinant DNA technology and chemical synthesis techniques. The application of peptides and proteins as clinically useful drugs is, however, seriously hampered owing to the substantial delivery problems requiring frequent injections. Considerable effort has been directed therefore to developing painless and convenient methods for delivery of peptides and proteins. In diabetes, in addition to the need of insulin injections there is a need to develop painless and convenient methods to measure blood glucose. This review describes approaches based on the application of ultrasound for noninvasive and painless transdermal glucose sensing and delivery of insulin.     

Lung surfactant: A biotechnological challenge

The genes for all three of the bona fide surfactant associated proteins have been cloned, allowing their production by recombinant DNA technology. In addition, improved protocols for the isolation of the natural surfactant proteins (NSP) made them available in larger quantities. Whereas, the NSP are often mixtures of allelic variants or functional isomers from gene families, the recombinant proteins (RSP) are obtained as single pure protein species. Antibodies directed against the N/RSP in combination with DNA probes have allowed new approaches to analyze the formation, location, transport, structure and functional capacities of these molecules as well as their interactions with one another and the phospholipids.     

体外培养的肝癌细胞株与正常肝细胞株蛋白质的差异表达

目的:运用SELDI蛋白质芯片技术分析体外培养的肝癌细胞株(HepG2)与正常肝细胞株(L02)蛋白质表达差异．方法:在体外培养HepG2和L02细胞株,收获细胞,将细胞用细胞裂解液裂解后,采用SELDI蛋白质芯片技术用IMAC3 及WCX2芯片检测HepG2、L02的蛋白质谱．结果:体外培养的肝癌细胞株与正常肝细胞株的蛋白质存在差异表达,IMAC3芯片共捕获61个蛋白,发现差异峰7个,与 L02细胞相比,其中3个差异蛋白在肝癌细胞中高表达,4个差异蛋白在肝癌细胞中低表达．WCX2芯片共捕获91个蛋白, 发现差异峰14个,其中3个差异蛋白在肝癌细胞中高表达,11 个差异蛋白在肝癌细胞中低表达．结论:SELDI蛋白芯片技术检测肝癌细胞株与正常肝细胞株蛋白质的差异表达方法简便,敏感性高,重复性好．