CONTRIBUTIONS OF MASS-SPECTROMETRY TO PEPTIDE AND PROTEIN-STRUCTURE
CONTRIBUTIONS OF MASS-SPECTROMETRY TO PEPTIDE AND PROTEIN-STRUCTURE
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DOI：
10.1002/bms.1200160119
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发表时间：
1988-10-01
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影响因子：
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通讯作者：
BIEMANN, K
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作者：
BIEMANN, K
随着重组脱氧核糖核酸（DNA）技术的出现，它允许对基因及其在细胞系中的表达进行广泛的操作，而不是在自然细胞系中，蛋白质结构的许多方面变得比以往任何时候都重要。除了确定与同源性有关的氨基酸序列问题，翻译后修饰的性质，由合成修饰基因产生的蛋白质结构的验证或自然突变体的检测，都是越来越频繁被问到的问题，并且需要更多的细节。质谱已成为该领域的重要贡献者，特别是自快速原子轰击（FAB）电离出现以来，这使得直接电离大极性分子（如肽和小蛋白质）成为可能。因此，FAB质谱法主要提供分子量信息，其本身通常足以回答某些问题，特别是因为许多肽的质量可以直接从混合物的单一质谱中确定。然而，为了获得详细的结构信息，例如氨基酸序列，必须通过碰撞过程诱导碎片化并分离产物离子，最好在串联系统的第二质谱仪中进行。这种方法特别适合于测定N -阻断肽的序列和阻断基团的性质；修饰（即磷酸化、硫酸化、糖基化）氨基酸的类型和位置；氨基酸替代的检测或验证；合成肽结构的确认；最后但同样重要的是，确定蛋白质的初级结构。
With the advent of recombinant deoxyribonucleic acid (DNA) technology which allows a wide range of manipulation of genes and their expression in cell lines other than the natural ones, many aspects of protein structure have become more important than ever. In addition to the determination of the amino acid sequence questions relating to homogeneity, the nature of post‐translational modifications, the verification of the structure of a protein produced by a synthetically modified gene or the detection of a natural mutant are all questions that are more and more frequently asked and with the demand for more detail. Mass spectrometry has emerged as an important contributor to this field, particularly since the advent of fast atom bombardment (FAB) ionization, which makes it possible to ionize directly large polar molecules such as peptides and small proteins. As such, FAB mass spectrometry provides mainly molecular weight information which, in itself, often sufficies to answer certain questions, particularly because the mass of many peptides can be determined directly from a single mass spectrum of a mixture. However, in order to obtain detailed structural information, such as the amino acid sequence, fragmentation has to be induced by collision processes and the product ions separated, preferably in the second mass spectrometer of a tandem system. This approach is particularly suited for the determination of the sequence of N‐blocked peptides and the nature of the blocking group; the type and location of modified (i.e. phosphorylated, sulfated, glycosylated) amino acids; detection or verification of amino acid replacements; confirmation of the structure of synthetic peptides; and last but not least, the determination of the primary structure of proteins.