糖复合物中糖链部分质谱测定

生物大分子结构与功能的研究是生物化学和分子生物学的一个核心内容,糖类的研究重新获得了人们的青睐,成为生命科学研究中继蛋白质、核酸后的下一个重要探索目标。十九世纪确定了碳水化合物的环状结构、立体构型和光学性质后,生物学家长期认为它除了提供能量维持生命外,只有结构组织的功能。五十年代蛋白质氨基酸组成的研究及后来的核酸高潮推动了生物化学和分子生物学的发展,七十年代免疫学的进展,认为抗体是一种含有共     

基于质谱的糖蛋白/糖链定量技术研究进展

糖基化修饰是一种重要的蛋白质翻译后修饰。糖基化修饰的蛋白质在生命体内具有重要的生物学功能。研究糖蛋白含量以及蛋白上糖链变化对于阐明糖基化修饰的功能具有重要的意义,也是当今的研究热点。本文就糖蛋白和糖链定量方法的研究进展和应用做了简要概述。     

糖组学研究中糖蛋白糖链结构分析技术

遗传信息由DNA传递至蛋白质,再经蛋白质翻译后糖基化修饰形成糖蛋白.与DNA、蛋白质相比,糖蛋白糖链结构更加多样,功能更加复杂,在一些重大的生理、病理事件中发挥着重要调节作用;而糖链如此复杂的功能是由其多样的结构决定的,糖链结构是糖组学研究的重要内容.本文就近年来糖组学研究中糖蛋白样品的提取分离、糖链释放及结构分析的基本方法及相关技术进展作了简要介绍.     

基于液相色谱-质谱联用技术的心肌缺血大鼠血清和心肌组织代谢组学

采用异丙肾上腺素诱导心肌缺血大鼠模型,使用液相色谱-质谱法检测血清和心肌中的内源性成分,应用软件对已鉴定的40余种目标成分进行靶向提取,用主成分分析(PCA)、有监督偏最小二乘法判别分析(PLS-DA)对代谢组学数据进行多维度统计分析,筛选潜在生物标志物。与对照组相比,在心肌缺血模型组大鼠血清、组织中检测出18个差异代谢物,涉及精氨酸和脯氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、谷氨酰胺和谷氨酸代谢、牛磺酸和亚牛磺酸代谢等多条代谢通路。代谢产物可作为心肌缺血研究中的重要标志物,该研究结果有助于揭示心肌缺血的发病机制,可为临床疾病诊断提供思路。     

基于高分辨质谱的石油组学分析技术的研究进展

随着原油重质化、劣质化程度的加深以及环境保护对燃油清洁性要求的日趋严格,对石油组成进行分子水平详细表征具有重大的现实意义和经济价值。依赖于高分辨质谱技术平台的石油组学分析技术成为实现这一目标的有力工具。本文以高分辨质谱技术平台为切入点,对石油组学定性定量分析技术的最新进展进行回顾和总结,并对其发展前景进行展望。     

基于质谱的蛋白质O-糖基化分析研究进展

蛋白质的O-糖基化是一种重要的蛋白质翻译后修饰,它和N-糖基化一样是蛋白质糖基化修饰的主要形式。蛋白质的O-糖基化对蛋白质的结构功能有重要的影响,因此分析蛋白质的O-糖基化具有重要的生物学意义。蛋白质O-糖基化分析包含4个方面的内容:(1)鉴定O-糖基化蛋白质的种类; (2)鉴定糖基化位点; (3)鉴定糖链结构; (4)糖链的定量分析。由于缺少保守的O-糖基化氨基酸特征序列,缺乏通用的糖苷酶以及O-糖链结构的复杂性等原因,基于质谱的蛋白质O-糖基化的分析目前仍处于方法开发阶段。本文主要介绍基于质谱的O-糖基化蛋白质的分析方法学在近期取得的一些进展,包括以下4个方面:O-糖蛋白/多肽的富集、O-糖链的解离、O-糖链的结构分析及O-糖基化定量分析。     

基于质谱技术的代谢组学研究及其在中国的发展

代谢组学是关于生物系统代谢物组成及变化规律的科学,是系统生物学的重要组成部分.质谱技术是目前代谢组学研究中最主要的分析手段之一,广泛应用于代谢组学各个领域.本文阐述了基于质谱技术的代谢组学方法及其应用,重点介绍和评论了近年来我国在该领域取得的进步和成果,并对基于质谱技术的代谢组学研究目前存在的问题及未来的发展进行了分析与展望.     

基于拟靶向液相色谱-质谱联用的胃癌患者血清代谢组分析

胃癌是一种高发的恶性肿瘤,是癌症相关死亡的第二大病因。早期筛查是提高患者生存率的有效手段,但目前临床上尚缺乏实现胃癌无创筛检的可靠标志物。本研究采用了基于液相色谱-质谱联用的拟靶向代谢组学方法分析了20例胃癌患者及40例正常人血清代谢组,以期发现新的潜在代谢标志物。代谢组数据的主成分分析和偏最小二乘法数据分析结果显示,胃癌患者与健康人群的血清代谢组存在明显的差异,结合非参数检验进一步筛选并定性出57个差异代谢物。其中二氢胆固醇经验证组样本验证,具有成为胃癌代谢标志物的潜力。本研究在发现胃癌的潜在代谢标志物的同时,也为胃癌患者代谢分型提供了重要的科学依据。     

糖环乙酰化苷类化合物的质谱研究

苷类化合物在自然界分布极为广泛,且有重要的生物活性。彭师奇等曾经比较系统地研究过的天麻苷和熊果苷,就有利尿、抗菌、降压等多方面的应用前景。糖和核酸及蛋白一样,是极其重要的内源性物质。研究糖衍生物的化学行为,对于揭示生命过程的重要现象有不容置疑的意义。本文选择天麻苷和熊果苷的乙酰化衍生物作为模型化合物,进行电子轰击谱(EI)、化学电离谱(CI)和快原子轰击谱(FAB)     

基于气相色谱-质谱联用技术的痛风病人血清代谢特征分析

痛风是一组仅见于人类的异质性疾病,随着时间的推移,将导致慢性关节炎并逐渐致残。该研究将基于气相色谱-质谱联用技术(GC-MS)的代谢组学方法应用于痛风病人的血清代谢特征分析。首先利用GCMS获得痛风病人和健康人的血清代谢指纹图谱,采用多变量统计分析对所得数据进行分析。主成分分析(PCA)得分图显示,痛风病人与健康人的血清代谢谱有差异。通过偏最小二乘-判别分析(PLS-DA)对样品进行进一步分型,根据模型的变量重要性因子(VIP值)及非参数检验结果筛选差异代谢物。共筛选出43种可能与痛风相关的代谢物,并对其中22个变量进行结构鉴定,主要包括丙二醇、2,3-二羟基丁酸、2,4-二羟基丁酸、赤藓糖醇、苏糖醇、苏糖酸、阿拉伯糖醇、D-葡萄糖酸、肌醇、次黄嘌呤、尿酸、尿苷、3-羟基-3-甲基丁酸、鸟氨酸、吲哚-3-乳酸、单乙醇胺、甘油、甘油酸、月桂酸及亚油酸等代谢物。与健康人相比,痛风病人的糖代谢、核苷酸代谢、氨基酸代谢及脂类代谢均发生了明显的紊乱。这些结果将为痛风临床诊断及治疗提供重要依据。     

Mass spectrometry analysis of melanoma related O-glycans in sera

Glycomics comparison was carried out by screening melanoma serum biomarkers between C57 mice with and without B16 implanted. O-glycans were released from 10 μL sera by β-elimination, purified by Graphitized Carbon Cartridge Solid Phase Extraction (GCC-SPE) and analyzed by MALDI-QIT-TOF-MS. MS raw data were acquired and exported by Launchpad software. MATLAB was then applied for further data analysis. 10 Glycans were considered to have stable changes after B16 implantation and 5 of them were under structural...     

Mass spectrometry in the structural analysis of flavonoids

Flavonoids are very common and widespread secondary plant metabolites. They have a wide range of biological and physiological activities and serve as chemotaxonomic marker compounds. Therefore, they have been extensively investigated both in the past and during recent years. The interest in them is still increasing. In the search for new compounds, and also in quality control, there is a need to have reliable methodology for the analysis of flavonoids. Mass spectrometry can make an invaluable contribution because of its high sensitivity, possibilities of coupling with liquid chromatography and the availability of powerful tandem mass spectrometric techniques. A review of currently available mass spectrometric methodology used in the structure elucidation of flavonoids is presented. Sample preparation, liquid chromatographic/mass spectrometric analysis and tandem mass spectrometric procedures for the characterization of flavonoid aglycones, O-glycosides, C-glycosides and acylated glycosides are considered.     

Mass spectrometry analysis of gliadins in celiac disease

In recent years, scientific research on wheat gluten proteins has followed three main directions aimed at (1) finding relationships between individual genetic alleles coding for gliadins, high or low molecular weight glutenin subunits, and the viscoelastic dough properties of flour-derived products such as pasta and bread; (2) identifying prolamins and derived peptides involved in celiac disease, a pathological condition in which the small intestine of genetically predisposed individuals is reversibly damaged; and (3) developing and validating sensitive and specific methods for detecting trace amounts of gluten proteins in gluten-free foods for celiac disease patients. In this review, the main aspects of current and perspective applications of mass spectrometry and proteomic technologies to the structural characterization of gliadins are presented, with focus on issues related to detection, identification, and quantification of intact gliadins, as well as gliadin-derived peptides relevant to the biochemical, immunological, and toxicological aspects of celiac disease.     

‘Extreme Mass Spectrometry’: the role of mass spectrometry in the study of the Antarctic Environment

A focus on the studies of the Antarctic environment that have been performed by mass spectrometry is presented herein; our aim is to give evidence of the essential role of this instrumental technique in the framework of the scientific research in Antarctica, with a comprehensive review on the main literature of the last two decades. Due to the wideness of the topic, the present review is limited to the determination of organic pollutants, natural molecules and biomarkers in Antarctica, thus excluding elemental analysis and studies on inorganic species. The work has been divided into five sections, on the basis of the considered environmental compartment: air; ice and snow; seawater, pack ice and lakes; soil and sediments; and organisms and biomarkers. Copyright © 2014 John Wiley & Sons, Ltd.     

Mass spectrometry in structural studies of diterpene alkaloids

Data on the use of electron impact mass spectrometry for the determination of the structures of diterpene alkaloids are surveyed and described systematically. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 6, pp. 1096–1104, June, 1997.     

Rapid and simple solid-phase esterification of sialic acid residues for quantitative glycomics by mass spectrometry

A rapid and quantitative method for solid-phase methyl esterification of carboxy groups of various sialylated oligosaccharides has been established. The method employed a triazene derivative, 3-methyl-1-p-tolyltriazene, for facile derivatization of oligosaccharides immobilized onto general solid supports such as Affi-Gel Hz and gold colloidal nanoparticles in a multiwell plate. The workflow protocol was optimized for the solid-phase processing of captured sialylated/unsialylated oligosaccharides separated from crude sample mixtures by chemical ligation. From tryptic and/or PNGase F-digest mixtures of glycoproteins, purification by chemoselective immobilization, esterification and recovery were achieved in the same well of the filter plate within three hours when used in conjunction with "glycoblotting technology" (S.-I. Nishimura, K. Niikura, M. Kurogochi, T. Matsushita, M. Fumoto, H. Hinou, R. Kamitani, H. Nakagawa, K. Deguchi, N. Miura, K. Monde, H. Kondo, High-throughput protein glycomics: Combined use of chemoselective glycoblotting and MALDI-TOF/TOF mass spectrometry: Angew. Chem. 2005, 117, 93-98; Angew. Chem. Int. Ed. 2005, 44, 91-96). The recovered materials were directly applicable to subsequent characterization by mass spectrometric techniques such as MALDI-TOF for large-scale glycomics of both neutral and sialylated oligosaccharides. On-bead/on-gold nanoparticle derivatization of glycans containing sialic acids allowed rapid and quantitative glycoform profiling by MALDI-TOF MS with reflector and positive ion mode. In addition to its simplicity and speed, the method eliminates the use of unfavorable halogenated solvents such as chloroform and dichloromethane or volatile solvents such as diethyl ether and hexane, resulting in a practical and green chemical method for automated robotic adaptation.     

Mass spectrometry of natural organic phosphorus

This article provides a review of the use of modern mass spectrometry (MS) for quantitative and qualitative measurements of organic phosphorus compounds in nature. Included is a brief discussion of recent developments in large molecule mass spectrometry, focusing on time-of-flight (TOF) and ion cyclotron resonance (ICR) mass analysis techniques, as well as electrospray (ESI) and inductively coupled plasma (ICP) ionization. The use of ICP with high-resolution mass spectrometry for quantitative measurements of total phosphorus and as a detector coupled to HPLC and CE for defining organic phosphorus speciation is demonstrated using results from a study of phosphorus cycling in a treatment wetland. Qualitative identifications of individual phosphorus compounds by ultrahigh resolution Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) is demonstrated using dissolved organic phosphorus isolated from this same wetland.     

Mass spectrometry of fluorocarbon‐labeled glycosphingolipids

A method for generation of novel fluorocarbon derivatives of glycosphingolipids (GSLs) with high affinity for fluorocarbon phases has been developed, and their potential applications to mass spectrometry (MS)‐based methodologies for glycosphingolipidomics have been investigated. Sphingolipid ceramide N‐deacylase (SCDase) is used to remove the fatty acid from the ceramide moiety, after which a fluorocarbon‐rich substituent (F‐Tag) is incorporated at the free amine of the sphingoid. In initial trials, a neutral GSL, globotriaosylceramide (Gb₃Cer), three purified bovine brain gangliosides, and four fungal glycosylinositol phosphorylceramides (GIPCs) were de‐N‐acylated, derivatized by prototype F‐Tags, and recovered by solid phase extraction on fluorocarbon‐derivatized silica (F‐SPE). The efficacy of SCDase treatment of GIPCs was here demonstrated for the first time. Compatibility with subsequent per‐N,O‐methylation was established for the F‐tagged Gb₃ Cer and purified gangliosides, and extensive mass spectra (MS¹ and MS²) consistent with all of the expected products were acquired. The potential use of F‐tagged derivatives for a comprehensive MS based profiling application was then demonstrated on a crude ganglioside mixture extracted from bovine brain. Finally, a simple trial in microarray format demonstrated fixation of F‐tagged G_M1 ganglioside to a fluorous glass surface, with the glycan intact and available for interaction with a fluorescent derivative of cholera toxin B chain. The methods described thus provide a new avenue for rapid GSL recovery or cleanup, potentially compatible with a variety of platforms for mass spectrometric profiling and structure analysis, as well as parallel analysis of functional interactions. Copyright © 2010 John Wiley & Sons, Ltd.     

Mass probe-assisted ionization method for total analysis of biomolecules with electrospray ionization-mass spectrometry

In this paper, we review the mass probes used for the derivation of a variety of biomolecules efficiently detected by the electrospray ionization-mass spectrometry and mass probe-assisted ionization method for total analysis and determination by consecutive detection with a single instrument. We describe mass probes for a variety of molecules including proteins, nucleobases, metallic cations, and other small molecules.     

基于质谱的蛋白质组学技术在神经退行性疾病生物标志物研究中的应用

蛋白质组学是在整体水平上研究细胞、组织或生物体蛋白质组成及变化规律的科学.与传统的生物学研究相比,蛋白质组学具有快速、灵敏、高通量的优点.神经退行性疾病是一类由神经系统内特定神经细胞的进程性病变或丢失而导致神经功能障碍的疾病,严重危害人类健康.近年来,基于质谱的蛋白质组学技术在神经退行性疾病的研究中得到了广泛应用.本文简要介绍了蛋白质组学在样品分离、多肽定量、质谱检测及生物标志物临床验证等方面的技术发展,并结合实例综述了基于质谱的蛋白质组学在神经退行性疾病生物标志物发现与验证中的研究进展.