RAPID IDENTIFICATION OF PROTEINS BY PEPTIDE-MASS FINGERPRINTING
RAPID IDENTIFICATION OF PROTEINS BY PEPTIDE-MASS FINGERPRINTING
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DOI：
10.1016/0960-9822(93)90195-t
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发表时间：
1993-06-01
期刊：
影响因子：
9.2
通讯作者：
BLEASBY, AJ
中科院分区：
文献类型：
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作者：
PAPPIN, DJC;HOJRUP, P;BLEASBY, AJ
背景：“软”电离技术的发展已经彻底改变了蛋白质结构分析的质谱方法。十多年来，这种技术已经被用于与特定蛋白酶的消化结合，以产生蛋白质的精确肽分子量“指纹”。这些指纹通常用于筛选已知的蛋白质，以检测翻译错误，表征翻译后修饰和分配二硫键。然而，在何种程度上肽质量信息可以单独使用，以确定未知的样品蛋白质，独立于其他分析方法，如蛋白质序列analysis.Results：我们在这里报告的分子量搜索（MOWSE）肽质量数据库的发展SERC达雷斯伯里实验室。实践经验表明，当针对来自超过50 000种蛋白质的片段数据库筛选这些肽质量时，可以从少至三个或四个实验确定的肽质量中唯一地鉴定样品蛋白质。几道尔顿的实验误差是容忍的评分算法，从而允许使用廉价的飞行时间质谱仪。与其他类型的物理数据，如氨基酸组成或线性序列，肽质量提供了一组的决定因素，是足够的歧视，以确定或匹配未知的样品protein.Conclusions：肽质量指纹可以证明作为歧视线性肽序列，但可以在一小部分的时间，使用较少的蛋白质。在许多情况下，这允许在将其提交给蛋白质序列分析之前快速鉴定样品蛋白质。片段质量还提供蛋白质水平的信息，与大规模DNA测序或绘图项目提供的信息互补。
Background: Developments in 'soft' ionisation techniques have revolutionized mass-spectrometric approaches for the analysis of protein structure. For more than a decade, such techniques have been used, in conjunction with digestion by specific proteases, to produce accurate peptide molecular weight 'fingerprints' of proteins. These fingerprints have commonly been used to screen known proteins, in order to detect errors of translation, to characterize post-translational modifications and to assign disulphide bonds. However, the extent to which peptide-mass information can be used alone to identify unknown sample proteins, independent of other analytical methods such as protein sequence analysis, has remained largely unexplored.Results: We report here on the development of the molecular weight search (MOWSE) peptide-mass database at the SERC Daresbury Laboratory. Practical experience has shown that sample proteins can be uniquely identified from as few as three or four experimentally determined peptide masses when these are screened against a fragment database that is derived from over 50 000 proteins. Experimental errors of a few Daltons are tolerated by the scoring algoridims, thus permitting the use of inexpensive time-of-flight mass spectrometers. As with other types of physical data, such as amino-acid composition or linear sequence, peptide masses provide a set of determinants that are sufficiently discriminating to identify or match unknown sample proteins.Conclusions: Peptide-mass fingerprints can prove as discriminating as linear peptide sequences, but can be obtained in a fraction of the time using less protein. In many cases, this allows for a rapid identification of a sample protein before committing it to protein sequence analysis. Fragment masses also provide information, at the protein level, that is complementary to the information provided by large-scale DNA sequencing or mapping projects.