Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometry
Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometry
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DOI：
10.1016/s1044-0305(03)00146-6
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发表时间：
2003-09-01
影响因子：
3.2
通讯作者：
Smith, RD
中科院分区：
文献类型：
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作者：
Strittmatter, EF;Ferguson, PL;Smith, RD
我们描述了应用毛细管液相色谱（LC）飞行时间（TOF）质谱仪器快速表征微生物蛋白质组。此前（Lipton et al.，Proc. Natl. Acad. Sci. U.S.A.2002，99，11049），已经使用毛细管LC MS/MS和精确质量测量表征了来自耐辐射异常球菌的一系列生长条件的肽，所述精确质量测量被捕获为精确质量和时间（AMT）标签数据库。使用该AMT标签数据库，由于额外使用洗脱时间数据作为约束，可以使用在TOF上获得的测量结果来分配检测到的肽。当使用AMT标签获得肽匹配（即，使用这两种约束）质谱峰的唯一匹配出现88%的时间。不仅AMT标签匹配在大多数情况下是独特的，蛋白质组的覆盖率也很高;在每个毛细管LC运行中平均发现了类似于3500个独特的肽AMT标签。从AMT标签数据库检索的结果来看，LC-TOFMS检测到了900个相似的ORF，其中500个相似的ORF被至少两个AMT标签覆盖。这些结果表明，具有适度质量和洗脱时间标准的AMT数据库搜索可以在单次运行中提供大约一千种蛋白质的蛋白质组学信息。
We describe the application of capillary liquid chromatography (LC) time-of-flight (TOF) mass spectrometric instrumentation for the rapid characterization of microbial proteomes. Previously (Lipton et al., Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 11049) the peptides from a series of growth conditions of Deinococcus radiodurans have been characterized using capillary LC MS/MS and accurate mass measurements which are captured as an accurate mass and time (AMT) tag database. Using this AMT tag database, detected peptides can be assigned using measurements obtained on a TOF due to the additional use of elution time data as a constraint. When peptide matches are obtained using AMT tags (i.e., using both constraints) unique matches of a mass spectral peak occurs 88% of the time. Not only are AMT tag matches unique in most cases, the coverage of the proteome is high; similar to3500 unique peptide AMT tags are found on average per capillary LC run. From the results of the AMT tag database search, similar to900 ORFs detected using LC-TOFMS, with similar to500 ORFs covered by at least two AMT tags. These results indicate that AMT database searches with modest mass and elution time criteria can provide proteomic information for approximately one thousand proteins in a single run of