Optimization of protein identification from digests as analyzed by capillary isoelectric focusing-mass spectrometry
Optimization of protein identification from digests as analyzed by capillary isoelectric focusing-mass spectrometry
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DOI：
10.1016/j.jchromb.2005.07.020
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发表时间：
2005-09-25
影响因子：
3
通讯作者：
van der Greef, J
中科院分区：
文献类型：
--
作者：
Storms, HF;van der Heijden, R;van der Greef, J
毛细管等电聚焦（CIEF）是一种用于分析多肽和蛋白质的高分辨率分离技术。当与离子阱质谱（CIEF-MS）结合时，独特的分离机制与高效的检测系统相结合。在较早的报告中，我们描述了与分析一组标准蛋白质的消化物有关的分离和接口方面。现在，我们报告蛋白质鉴定过程的不同方面。Sequest软件参数通过使用标准蛋白质消化物进行优化。这些设置用于分析大肠杆菌的周质蛋白。由于在CIEF中，肽根据其pI值集中，因此特定肽的动员时间取决于其pI值。基于这种关系，一些肽的鉴定是方便的。此外，可以评估所使用的Sequest设置。总共在一次运行中鉴定了159种蛋白质。(c)2005 Elsevier B.V.保留所有权利。
Capillary isoelectric focusing (CIEF) is a high-resolution separation technique for the analysis of peptides and protein digests. When coupled to ion trap-mass spectrometry (CIEF-MS) the unique separation mechanism is combined with a highly efficient detection system. In an earlier report, we described aspects of separation and interfacing in connection to the analysis of a digest of set of standard proteins. Now, we report on different aspects of the process of protein identification. Sequest software parameters were optimized by using a standard protein digest. These settings were used for the analysis of periplasmic proteins from Escherichia coh. Since in CIEF peptides are focused according to their pI values, the mobilization time of a particular peptide is dependent on its pI value. Based on this relation, the identification of some peptides was facilitated. Furthermore, the Sequest settings that were used could be evaluated. In total, 159 proteins were identified in a single run. (c) 2005 Elsevier B.V. All rights reserved.