表面增强激光解吸电离质谱芯片在蛋白质组学研究中的应用

蛋白质组学是当今生命科学研究中最活跃的前沿领域。其主要技术手段是双向凝胶电泳,质谱技术和生物信息学,但各有其局限性。芯片技术和质谱技术相结合产生的表面增强激光解析离子化－飞行时间－质谱9SELDI－TOF－MS）技术,将蛋白质样品的制备,生化反应到检测分析的整个过程集成在芯片上进行,实现了新型,高效,快速,高通量的检测,不仅促进了蛋白质组学研究的迅速发展,而且为阐明生命活动的规律和疾病的发病机理以及寻找合适的医药提供提供了有力的工具。     

生物质谱与蛋白质组学

蛋白质组学是后基因组学时代最受关注的研究领域之一,其核心的鉴定技术——生物质谱近年来在仪器设计以及鉴定通量、分辨率和灵敏度等各方面均有质的飞跃,促进了蛋白质表达谱作图、定量蛋白质组分析、亚细胞器蛋白质组作图、蛋白质翻译后修饰以及蛋白质相互作用等蛋白质组研究各个领域的飞速发展。本综述了生物质谱技术的最新进展,及其在蛋白质组学研究中的应用。     

LCM纯化的人支气管上皮癌变各阶段组织的定量蛋白质组学研究

为分析支气管上皮癌变进程中的差异表达蛋白质,筛选肺鳞癌早期诊断标志物,以人支气管上皮癌变各阶段组织为研究对象,先采用激光捕获显微切割技术(LCM) 纯化人正常支气管上皮组织、鳞状化生、不典型增生、原位癌、浸润性肺鳞癌组织,再用同位素标记相对和绝对定量 (iTRAQ) 技术结合二维液相色谱串联质谱（2D LC-MS/MS）鉴定支气管上皮癌变进程中各阶段的差异表达蛋白质。结果共鉴定了1036个蛋白质,筛选出102个与人支气管上皮癌变相关的差异蛋白质,在这些差异蛋白质中,有的在支气管上皮癌变过程中进行性上调,有的在支气管上皮癌变过程中进行性下调,有的呈阶段特异性改变;功能分析表明,这些差异蛋白质涉及代谢、细胞凋亡、增殖、分化、信号传导、转录、翻译、细胞粘附、免疫反应与发育等。Western blotting 及免疫组织化学技术验证了其中 2个差异蛋白(S100A9和 CKB) 的表达,证实了定量蛋白质组学结果的可靠性。研究结果提示：这些差异表达蛋白质与支气管上皮癌变相关,并可成为肺鳞癌的早期诊断标志物,进一步研究差异蛋白的生物学功能,将有助于阐明支气管上皮的癌变机制,从而为肺鳞癌的早期诊断与发病机制研究提供新思路。     

蛋白质组学研究技术及其联合应用

蛋白质组学是研究细胞、组织和器官内所有蛋白质的组成及其动态变化的科学,是在蛋白质水平上定量的、动态的、整体的研究生物体。目前蛋白质组学技术分为样品制备、分离和鉴定3个方面,其新技术主要有激光捕捉显微解剖法、离心超滤法、双向凝胶电泳、同位素亲和标签技术、色谱技术以及质谱技术等。然而,任何一种蛋白质组学研究技术都有其缺陷。因此多种技术的联合应用能使蛋白质组研究更精确和完整,是蛋白质组学的发展趋势。     

植物蛋白质组学研究若干重要进展

植物蛋白质组学近年来正从定性向精确定量蛋白质组学的方向发展。国际上近两年发表的约160篇研究论文报道了利用不断改进的双向电泳结合生物质谱技术、多维蛋白质鉴定技术,以及包括双向荧光差异凝胶电泳、幅N体内代谢标记、同位素标记的亲和标签、同位素标记相对和绝对定量等在内的第2代蛋白质组学技术,对植物组织（器官）与细胞器、植物发育过程和植物响应环境胁迫的蛋白质组特征,以及植物蛋白质翻译后修饰和蛋白质相互作用等方面的研究成果。该文对上述报道进行总结,综述了2007年以来植物蛋白质组学若干重要问题研究的新进展。     

胆管癌患者与健康志愿者血清样品蛋白质表达差异分析

胆管癌（cholangiocarcinoma,CCA）是一种难于早期诊断和治疗的恶性肿瘤.利用蛋白质组学技术以期筛选出人血清中具有医学价值的胆管癌癌变的标志物.利用白蛋白/IgG去除试剂盒去除血清中高丰度的白蛋白和IgG,再经丙酮沉淀得到高质量的血清样品.样品经双向电泳（2-DE）得到了分辨率较好的胆管癌血清的蛋白点图谱.利用Image Master 2D软件对比分析了胆管癌病人和正常健康对照组的2-DE图谱.其中,表达上调蛋白有8个,表达下调蛋白有6个.用质谱获得上述差异蛋白的肽质量指纹图谱,并经数据库检索共鉴定出了11种差 异蛋白质,其中8种蛋白质的表达与胆管癌密切相关.本研究为阐明胆管癌的机理提供了一条新途径.     

串联质谱数据从头解析与蛋白质的数据库搜索鉴定

蛋白质的鉴定是蛋白质组学研究中必不可少的一步。用串联质谱（ｔａｎｄｅｍ ｍａｓｓ ｓｐｅｃｔｒｏｍｅｔｒｙ,ＭＳ／ＭＳ）可以进行多肽的从头测序（ｄｅ ｎｏｖｏ ｓｅｑｕｅｎｃｉｎｇ）,并搜索数据库以鉴定蛋白质。用图论以及真实谱－理论谱联配（ａｌｉｎｇｍｅｎｔ）的方法对串联质谱得到的多肽图谱进行从头解析,得到了可靠的多肽序列,并应用到数据库搜索中鉴定了相应的蛋白质。同时,还用统计的方法对ＳｗｉｓｓＰ     

蛋白质组学在结核分枝杆菌研究中的应用

蛋白质组学是在基因组学基础上发展起来的新兴学科, 其基本技术包括样品制备、蛋白质分离和蛋白质鉴定分析, 其中的核心技术是双向凝胶电泳技术(2-Dimensional Electrophoresis, 2-DE)和质谱技术(Mass Spectrometry, MS)。近年来, 蛋白质组学技术已应用于结核分枝杆菌的研究领域。应用蛋白质组学技术分离、鉴定、检测结核分枝杆菌致病株的全菌蛋白及分泌蛋白, 分析其蛋白组成, 可深入解析结核分枝杆菌的致病机理和耐药机制。通过对结核分枝杆菌致病株抗原的分析, 为研制预防结核病的新型疫苗拓展了空间。通过对结核分枝杆菌临床分离株的蛋白组成分析还发现了一些有意义的结核病早期诊断标志物。蛋白质组学技术还应用于寻找新的药物靶标, 在研制和筛选新的抗结核药物等方面展示了一些有价值的研究成果, 为更好地开展结核病的预防、早期诊断及治疗打下了基础。     

水稻蛋白质组学研究进展

蛋白质组是一个基因组所表达蛋白质的总称,研究内容包括蛋白质基本氨基酸序列的鉴定到相关性状、修饰、功能及不同类型蛋白分子相互作用等等。本文简要介绍了蛋白质组学的产生背景,以及主要研究技术包括双向电泳(2D-PAGE)质谱、蛋白质芯片、酵母双杂交,并重点介绍了近期水稻蛋白质组学应用研究进展。     

质谱兼容的蛋白质银染色法

蛋白质组学是对蛋白质组进行研究的一门新兴学科。它可以揭示细胞内蛋白质组成成分与修饰状态的动态变化．研究蛋白质之间的相互作用．从全局的高度来研究代谢．发育以及调控等复杂的问题。 由于蛋白质组学研究范围十分广泛，所需要的技术手段也多种多样，如双向凝胶电泳（2-DE）．色谱．蛋白质芯片、质谱以及生物信息学等。2-DE分离蛋白质．通过生物质谱以及生物信息学手段对蛋白质进行鉴定是目前最常用的一种研究策略。由于该套技术平台中影响因素较多．很多初学者难以快速掌握．所以本实验方法系列讲座将详细介绍这一常规技术平台中的多项经典实验方案（样品制备．2一DE、染色、质谱鉴定等）．并着重对一些常见问题与难点进行深入的探讨。此外，本实验方法系列讲座还将涉及近年来刚刚兴起的大规模的蛋白质组自动分析系统——液相色谱-质谱联用．旨在为研究者拓宽实验方法的视野。[编者按]     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.