质谱与生命科学

本文以实验表明了谱学，特别是快原子轰击质谱（ＦＡＢ－ＭＳ）、电喷雾电离质谱（ＥＳＩ－ＭＳ）、串联质谱（ＭＳ－ＭＳ）以及有底物的激光解吸电离飞行时间质谱（ＭＡＬＤＩ－ＴＯＦ－ＭＳ）等技术在解决生物物质－肽－蛋白质等的一级结构难题中的特定作用，而这些结构难题往往是常规的生物不方法－Ｅｄｍａｎ降解法、ＤＮＡ转泽法等所无能为力的。虽然文中讨论的例子仅涉及到肽、蛋白质的结构修饰，同理，它也可以广泛应用于其他     

毛细管电泳在形态分析中的应用

对近年来毛细管电泳（ＣＥ）与紫外检测器（ＵＶｄｅｔｅｃｔｏｒ）和ＣＥ与感应耦合等离子体质谱（ＩＣＰ－ＭＳ）联用在形态分析中的应用及其存在的一些问题加以评述。     

MALDI－TOF－MS法分析人血清白蛋白

用基质辅助激光解吸附飞行时间质谱（ＭＡＬＤＩ－ＴＯＦ－ＭＳ）法分析测定了人血清白蛋白。结果表明，全军血液制品研究中心应用Ｒｉｖａｎｏｌ和Ｃｏｈｎ＇ｓ两种方法分离提取的人血清白蛋白，其分子量与理论计算值相吻合，其纯度与市售标准人血清白蛋白相一致，用ＭＡＬＤＩ－ＴＯＦ－ＭＳ法分析鉴定人血清白蛋白，准确、简便和灵敏（仅需ＰＭ浓度样品），且重复性好，日内和日间差的变异系数均小于０．０３％，是ＳＤＳ－ＰＡＧＥ电泳法所无法比拟的。     

芳香硝基化合物的能量分辨质谱

本文报道用ＭＳ／ＭＳ技术的一种重要实验方法——能量分辨质谱（ＥＲＭＳ）来鉴别质谱分析中的同分异构体化合物。系统探讨了同分异构的硝基芳香化合物２，３－ＤＮＴ和３，５－ＤＮＴ（相对分子质量为１８２）在电子轰击（ＥＩ）方式下，裂分途径与其分子内能的相互关系以及离子的结构和裂分途径相对于碰撞能量（ＣＥ）的依赖关系（裂分曲线）。结果表明，利用同分异构化合物在能量分辨中的显著差异可以辨别它们的结构     

毛细管电泳质谱联用技术及其应用

本文介绍了用于毛细管电泳质谱联用仪器的多种接口技术，描述了ＣＺＥ，ＣＩＥＦ，ＣＧＥ，ＭＥＫＣ和ＣＩＴＰ等毛细管电泳技术和四极质谱，离子阱质谱，傅 叶变换离子回旋共振质谱，飞行时间质谱，磁质谱，解吸质谱等联用的现状及发展前景，对近年来ＣＥ－ＭＳ在酶解产物。蛋白质和肽，核苷酸，药物及代谢产物等领域中的应用作了详细述评。     

有机锡化合物的电喷雾电离质谱

报道了一系列有机锡ＲＲＰ（Ｓ）ＳＳｎ（Ｃｙ）３化合物的电喷雾电离质谱（ＥＳＩ－ＭＳ）及碰撞诱导解离质谱（ＣＩＤ－ＭＳ）,总结了其谱图特征,讨论了其碎裂机理,为该类化合物的鉴定提供了依据。     

等离子体原子发射光谱/质谱在新型陶瓷材料分析中的应用

本文对近几年来等离子体原子发射光谱（ＩＣＰ－ＡＥＳ）和等离子体质谱（ＩＣＰ－ＭＳ）在新型陶瓷材料分析中的应用进行了评述,讨论了应用ＩＣＰ－ＡＥＳ／ＭＳ技术测定陶瓷材料中痕量杂质的各种进样技术。引用文献６０篇。     

电喷雾电离质谱在富勒烯化学中的应用进展

电喷才电离质谱（ＥＳＩ－ＭＳ）是本世纪发展的一种非常重要的质谱，具有软电离、无碎片的特点，可用于分析检测非挥发性的，极性的、热不稳定的化合物，在富勒烯化学研究中起了重要作用。本文对ＥＳＩ－ＭＳ在富勒烯化学中的应用进展作一概要评述。     

苯并硫杂冠醚的质谱研究

报道了两个苯并硫杂冠醚的电子电离质谱，利用高分辩质谱（ＨＲＭＳ）和碰撞活化－质量分析动能谱（ＣＡＤ－ＭＩＫＥ谱）研究了它们的离子碎裂途径。苯并硫杂冠醚含两个硫原子的碎片离子进一步断裂时以丢失Ｃ２Ｈ４Ｓ为特征。     

快原子轰击和电喷雾电离质谱表征烷基苯磺酸盐

用负离子化快原子轰击（ＦＡＢ）和电喷雾电离（ＥＳＩ）质谱并结合碰撞活化解离（ＣＡＤ）质谱方法对烷基苯磺酸盐（ＡＢＳ）进行鉴定。试样在负离子电喷雾电离（ＥＳＩ）过程中不产生碎片峰,但是在ＦＡＢ过程中产生许多碎片、比较这些谱图,就可以区别碎片峰和分子离子峰。线性或支化烷基苯磺酸盐的结构与相对含量可以在离子ＥＳＩ／ＣＡＤ－ＭＳ方法中得到。负离子ＥＳＩ－ＭＳ是快速、有效和可靠的鉴定ＡＢＳ的方法。     

人的基因组及DNA序列分析──过去、现去及将来（上）

人的基因组研究已成为生命科学前沿领域中最热门的课题之一。ＤＮＡ序列分析是基因组研究的关键技术．本文对人的基因组分析及其对ＤＮＡ序列分析的要求进行了论述．对ＤＮＡ序列分析方法如板凝胶电泳自放射显影法、板凝胶电泳激光荧光法、毛细管电泳激光荧光法、阵列毛细管凝胶电泳激光荧光法。超薄层板在胶电泳激光荧光法作了详细评论．并对正在开发的不用凝胶电泳分离的直接测序新技术和新方法，如质谱法、原子探针法（扫描隧道显微镜、原子力显微镜）、杂交法、流动单分子荧光检测法等进行了评论。     

DNA sequencing

Determination of the sequence of DNA is one of the most important aspects of modern molecular biology. New sequencing methods currently being developed enable DNA sequence to be determined increasingly faster and more efficiently. One of the major advances in sequencing technology is the development of automated DNA sequencers. These utilize fluorescent rather than radioactive labels. A laser beam excites the fluorescent dyes, the emitted fluorescence is collected by detectors, and the information analyzed by computer. Robotic work stations are being developed to perform template preparation and purification, and the sequencing reactions themselves. Research is currently in progress to develop the technology of mass spectrometry for DNA sequencing. Success in this endeavor would mean that the gel electrophoresis step in DNA sequencing could be eliminated. A major innovation has been the application of polymerase chain reaction (PCR) technology to DNA sequence determination, which has led to the development of linear amplification sequencing (cycle sequencing). This very powerful yet technically simple method of sequencing has many advantages over conventional techniques, and may be used in manual or automated methods. Other recent innovations proposed recently to increase speed and efficiency include multiplex sequencing. This consists of pooling a number of samples and processing them as pools. After electrophoresis, the DNA is transferred to a membrane, and sequence images of the individual samples are obtained by sequential hybridizations with specific labeled oligonucleotides. Multiplex DNA sequencing has been used in conjunction with direct blotting electrophoresis to facilitate transfer of the DNA to a membrane. Chemiluminescent detection can also be used in conjunction with multiplex DNA sequencing to visualize the image on the membrane.     

蛋白质顺序测定的策略

近年来蛋白质顺序测定在自动化、微量化与策略方面都有显著的进展。用DNA顺序分析技术间接地测定蛋白质顺序的策略已被广泛地采用,它能加快但不能代替蛋白质的顺序测定。质谱技术测定蛋白质顺序的途径也有了可喜的突破。高效液相色谱分离肽技术的发展、限制性酶切及各种选择性的化学与酶的降解方法的出现,为蛋白质顺序测定提供了更多可选用的工具。     

Tandem mass spectrometry methods for definitive protein identification in proteomics research

Optimized procedures have been developed for the addition of sulfonic acid groups to the N-termini of low-level peptides. These procedures have been applied to peptides produced by tryptic digestion of proteins that have been separated by two-dimensional (2-D) gel electrophoresis. The derivatized peptides were sequenced using matrix-assisted laser desorption/ionization (MALDI) post-source decay (PSD) and electrospray ionization-tandem mass spectrometry methods. Reliable PSD sequencing results have been obtained starting with sub-picomole quantities of protein. We estimate that the current PSD sequencing limit is about 300 fmol of protein in the gel. The PSD mass spectra of the derivatized peptides usually allow much more specific protein sequence database searches than those obtained without derivatization. We also report initial automated electrospray ionization-tandem mass spectrometry sequencing of these novel peptide derivatives. Both types of tandem mass spectra provide predictable fragmentation patterns for arginine-terminated peptides. The spectra are easily interpreted de novo, and they facilitate error-tolerant identification of proteins whose sequences have been entered into databases.     

Capillary electrophoresis of proteins for proteomic studies

Analyses of proteins in complex mixtures such as cell lyzates are presently performed mainly by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. For structural analysis, each protein in a spot is digested with proteases and the fragment peptides are subjected to Edman sequencing and/or mass spectrometry. These works aim at the total analysis of proteins in a complex mixture and reconstruction of their cooperative functions. Genomic studies are now being combined with these proteomic studies. This review article focuses on the application of capillary electrophoresis aiming at the total analysis of complex protein systems or structural analysis of each separated protein. From this viewpoint, articles on capillary zone electrophoresis, capillary isoelectric focusing, and sieving SDS capillary electrophoresis are reviewed. Since these techniques of capillary electrophoresis have been thoroughly reviewed previously, papers published in 1997 and 1998 are mainly covered.     

质谱及相关技术在信号传导方面的研究进展

细胞信号传导是近年来生命科学研究的热点之一。有关蛋白转录后修饰 (如蛋白质磷酸化、乙酰化、糖基化等 ) ,信号肽序列测定 ,信号传导途径和多通道调节方式 ,蛋白自折叠及构象变化 ,小分子脂类信号分子等研究由于质谱技术的快速发展而取得了突破性的进展     

一种基于肽质量指纹谱技术鉴定蓖麻毒素的新方法

应用基质辅助激光解吸电离飞行时间质谱(MALD I-TOF/MS)法实现了对蓖麻毒素(R ic in)的鉴定。测定蓖麻毒素的分子量为62925Da,实现了蓖麻粗毒的凝胶电泳分离,并通过胶内酶切获得了蓖麻毒素的肽质量指纹谱(PMF);经过数据库检索,在输入检索的22条肽段中有17条获得了匹配。检索结果显示,利用生物质谱技术是鉴定蓖麻毒素的最有效的新方法之一。     

Mass spectrometric identification of proteins and characterization of their post-translational modifications in proteome analysis

High-throughput DNA sequencing has resulted in increasing input in protein sequence databases. Today more than 20 genomes have been sequenced and many more will be completed in the near future, including the largest of them all, the human genome. Presently, sequence databases contain entries for more than 425.000 protein sequences. However, the cellular functions are determined by the set of proteins expressed in the cell--the proteome. Two-dimensional gel electrophoresis, mass spectrometry and bioinformatics have become important tools in correlating the proteome with the genome. The current dominant strategies for identification of proteins from gels based on peptide mass spectrometric fingerprinting and partial sequencing by mass spectrometry are described. After identification of the proteins the next challenge in proteome analysis is characterization of their post-translational modifications. The general problems associated with characterization of these directly from gel separated proteins are described and the current state of art for the determination of phosphorylation, glycosylation and proteolytic processing is illustrated.     

Membrane preconcentration-capillary electrophoresis tandem mass spectrometry (mPC-CE-MS/MS) in the sequence analysis of biologically derived peptides

We demonstrate the use of membrane preconcentration capillary electrophoresis tandem mass spectrometry (mPC-CE-MS/MS) for sequencing peptides at the sub-100 femtomole level. In particular by loading the mPC-CE cartridge off-line with a pressurized bomb apparatus, 100 mul solutions can be loaded in <5 min. Furthermore, mPC-CE-MS in conjunction with on-line transient isotachophoresis carried out in 25 mum i.d. capillaries results in enhanced resolution and theoretical plate values as compared to convention 50-75 mum i.d. capillaries. We show that this is a powerful new approach in the sequencing of biologically derived compounds from complex mixtures such as MHC class I peptides.     

复杂生物体系中蛋白质高效分离分析技术的新进展

继人类基因组计划完成之后,作为一种新的研究策略,蛋白质组学在生命科学研究中发挥着愈来愈重要的作用。由于生物体系的复杂性和多样性,使得分离效率高、灵敏度高、通量高和动态范围宽的分离分析技术平台的研究和应用已成为蛋白质组学研究的重点和热点之一。着重介绍了近年来应用日益广泛的多维色谱预分离、毛细管液相色谱-质谱联用、毛细管电泳及其与质谱联用等高效分离分析技术在复杂生物体系的蛋白质分析中的最新进展。引用相关文献40篇。