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研究肽段从头测序需要解决的问题之一是区分亮氨酸和异亮氨酸残基,利用基质辅助激光解吸电离-串联飞行时间质谱(MALDI-TOF/TOF)所产生的w型离子区分这两种氨基酸残基是目前最常用的方法。本研究以6对合成的同分异构肽段（由7~15个氨基酸构成）为例,来说明w离子信号强度较低且有时不能产生,在区分多肽中的亮氨酸和异亮氨酸时受肽段长度和序列的限制。而亮氨酸与异亮氨酸的变化会导致亚胺相关离子、内部断裂离子、b-型和y-型离子等相对强度的微小变化。通过比较这些骨架断裂变化所产生的微小差异,可为区分肽段中的亮氨酸和异亮氨酸残基提供新的线索和判据。 相似文献
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Albert T. Lebedev Irina D. Vasileva Tatiana Y. Samgina 《Mass spectrometry reviews》2022,41(2):284-313
The present review covers available results on the application of FT-MS for the de novo sequencing of natural peptides of various animals: cones, bees, snakes, amphibians, scorpions, and so forth. As these peptides are usually bioactive, the animals efficiently use them as a weapon against microorganisms or higher animals including predators. These peptides represent definite interest as drugs of future generations since the mechanism of their activity is completely different in comparison with that of the modern antibiotics. Utilization of those peptides as antibiotics can eliminate the problem of the bacterial resistance development. Sequence elucidation of these bioactive peptides becomes even more challenging when the species genome is not available and little is known about the protein origin and other properties of those peptides in the study. De novo sequencing may be the only option to obtain sequence information. The benefits of FT-MS for the top-down peptide sequencing, the general approaches of the de novxxo sequencing, the difficult cases involving sequence coverage, isobaric and isomeric amino acids, cyclization of short peptides, the presence of posttranslational modifications will be discussed in the review. 相似文献
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Amino acid-tagging strategies are widespread in proteomics. Because of the central role of mass spectrometry (MS) as a detection technique in protein sciences, the term \"mass tagging\" was coined to describe the attachment of a label, which serves MS analysis and/or adds analytical value to the measurements. These so-called mass tags can be used for separation, enrichment, detection, and quantitation of peptides and proteins. In this context, cysteine is a frequent target for modifications because the thiol function can react specifically by nucleophilic substitution or addition. Furthermore, cysteines present natural modifications of biological importance and a low occurrence in the proteome that justify the development of strategies to specifically target them in peptides or proteins. In the present review, the mass-tagging methods directed to cysteine residues are comprehensively discussed, and the advantages and drawbacks of these strategies are addressed. Some concrete applications are given to underline the relevance of cysteine-tagging techniques for MS-based proteomics. 相似文献
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两种蜘蛛多肽毒素的电喷雾电离质谱法序列分析 总被引：1，自引：0，他引：1
虎纹捕鸟蛛毒素-(HWTX-)和敬钊缨毛蛛毒素-(JZTX-)是两种蜘蛛多肽类神经毒素。它们的大部分氨基酸序列已由Edman降解法测出,但仍有部分序列由于某些原因未能确定。为了鉴定未知部分的序列,利用电喷雾电离-四极杆-飞行时间质谱(ESI-Qq-TOF-MS)对两种蜘蛛毒素的酶解产物进行分析,获得其酶解片段的一级(MS)和二级(MS/MS)图谱,在用软件和手工的方式对含有未知序列的肽段进行解析后,得到未知部分确切氨基酸序列。研究表明:利用质谱的从头测序(denovosequencing)方法获得HWTX-和JZTX-完整的氨基酸序列。该方法在蛋白质和多肽的结构分析和鉴定与传统方法相比具有高灵敏度和快速的优势,是对传统Edman化学降解法的一个很好的补充。 相似文献
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The role of mass spectrometry in plant systems biology 总被引：10，自引：0，他引：10
Large-scale analyses of proteins and metabolites are intimately bound to advancements in MS technologies. The aim of these non-targeted "omic" technologies is to extend our understanding beyond the analysis of only parts of the system. Here, metabolomics and proteomics emerged in parallel with the development of novel mass analyzers and hyphenated techniques such as gas chromatography coupled to time-of-flight mass spectrometry (GC-TOF-MS) and multidimensional liquid chromatography coupled to mass spectrometry (LC-MS). The analysis of (i) proteins (ii) phosphoproteins, and (iii) metabolites is discussed in the context of plant physiology and environment and with a focus on novel method developments. Recently published studies measuring dynamic (quantitative) behavior at these levels are summarized; for these works, the completely sequenced plants Arabidopsis thaliana and Oryza sativa (rice) have been the primary models of choice. Particular emphasis is given to key physiological processes such as metabolism, development, stress, and defense. Moreover, attempts to combine spatial, tissue-specific resolution with systematic profiling are described. Finally, we summarize the initial steps to characterize the molecular plant phenotype as a corollary of environment and genotype. 相似文献
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Protein post‐translational modifications (PTMs) are critically important in regulating both protein structure and function, often in a rapid and reversible manner. Due to its sensitivity and vast applicability, mass spectrometry (MS) has become the technique of choice for analyzing PTMs. Whilst the “bottom‐up' analytical approach, in which proteins are proteolyzed generating peptides for analysis by MS, is routinely applied and offers some advantages in terms of ease of analysis and lower limit of detection, “top‐down” MS, describing the analysis of intact proteins, yields unique and highly valuable information on the connectivity and therefore combinatorial effect of multiple PTMs in the same polypeptide chain. In this review, the state of the art in top‐down MS will be discussed, covering the main instrumental platforms and ion activation techniques. Moreover, the way that this approach can be used to gain insights on the combinatorial effect of multiple post‐translational modifications and how this information can assist in studying physiologically relevant systems at the molecular level will also be addressed. © 2012 Wiley Periodicals, Inc., Mass Spec Rev 32:27–42, 2013 相似文献
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Wenna Nie Leyu Yan Yie H. Lee Chandan Guha Irwin J. Kurland Haitao Lu 《Mass spectrometry reviews》2016,35(3):331-349
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Elien Vandermarliere Elisabeth Stes Kris Gevaert Lennart Martens 《Mass spectrometry reviews》2016,35(6):653-665
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Ganesh Kumar Agrawal Abhijit Sarkar Pier Giorgio Righetti Romina Pedreschi Sebastien Carpentier Tai Wang Bronwyn J. Barkla Ajay Kohli Bongani Kaiser Ndimba Natalia V. Bykova Christof Rampitsch Lello Zolla Mohamed Suhail Rafudeen Rainer Cramer Laurence Veronique Bindschedler Nikolaos Tsakirpaloglou Roya Janeen Ndimba Jill M. Farrant Jenny Renaut Dominique Job Shoshi Kikuchi Randeep Rakwal 《Mass spectrometry reviews》2013,32(5):335-365
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Recent literature that highlights the power of using mass spectrometry (MS) for protein identification from preparations of highly purified organelles and other large subcellular structures is covered in this review with an emphasis on techniques that preserve the integrity of the functional protein complexes. Recent advances in distinguishing contaminant proteins from "bonafide" organelle-localized proteins and the affinity capture of protein complexes are reviewed, as well as bioinformatic strategies to predict protein organellar localization and to integrate protein-protein interaction maps obtained from MS-affinity capture methods with data obtained from other techniques. Those developments demonstrate that a revolution in cellular biology, fueled by technical advances in MS-based proteomic techniques, is well underway. 相似文献
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Dityrosine is a fluorescent molecule formed as a result of normal posttranslational processing. In many structural proteins, dityrosine confers resistance to proteolysis and physicochemical trauma as a stabilizing crosslink. Dityrosine has also been found in oxidative/nitrative stress under a variety of conditions and biological systems. In this regard, it has been used as an important biomarker for oxidatively modified proteins during UV and gamma-irradiation, aging, and exposure to oxygen free radicals, nitrogen dioxide, peroxynitrite, and lipid hydroperoxides. Renewed interest in dityrosine and other tyrosine oxidation products as clinical indicators of oxidative modification has driven the development of important techniques for the specific analysis and quantification of these molecules. The presence of elevated levels of dityrosine in mammalian tissue and urine samples has been measured by chromatographic separation followed by mass spectrometry GC-MS and HPLC-MS/MS. Increases in dityrosine levels have been associated with pathologies such as eye cataracts, atherosclerosis, acute inflammation, and Alzheimer's disease. The continued development of, and increased accessibility to, improved mass spectrometric instrumentation will expand the capability, feasibility, and sensitivity with which specific biomarkers like dityrosine can be measured. 相似文献