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首页> 外文期刊>Journal of proteome research >Comparison of vacuum matrix-assisted laser desorption/ionization (MALDI) and atmospheric pressure MALDI (AP-MALDI) tandem mass spectrometry of 2-dimensional separated and trypsin-digested glomerular proteins for database search derived identification
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Comparison of vacuum matrix-assisted laser desorption/ionization (MALDI) and atmospheric pressure MALDI (AP-MALDI) tandem mass spectrometry of 2-dimensional separated and trypsin-digested glomerular proteins for database search derived identification

机译：二维分离和胰蛋白酶消化的肾小球蛋白的真空基质辅助激光解吸/电离（MALDI）和大气压MALDI（AP-MALDI）串联质谱的比较，用于数据库搜索衍生的鉴定

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Mass spectrometric based sequencing of enzymatic generated peptides is widely used to obtain specific sequence tags allowing the unambiguous identification of proteins. In the present study, two types of desorption/ionization techniques combined with different modes of ion dissociation, namely vacuum matrix-assisted laser desorption/ionization (vMALDI) high energy collision induced dissociation ( CID) and post-source decay (PSD) as well as atmospheric pressure (AP)-MALDI low energy CID, were applied for the fragmentation of singly protonated peptide ions, which were derived from two-dimensional separated, silver-stained and trypsin-digested hydrophilic as well as hydrophobic glomerular proteins. Thereby, defined properties of the individual fragmentation pattern generated by the specified modes could be observed. Furthermore, the compatibility of the varying PSD and CID (MS/MS) data with database search derived identification using two public accessible search algorithms has been evaluated. The peptide sequence tag information obtained by PSD and high energy CID enabled in the majority of cases an unambiguous identification. In contrast, part of the data obtained by low energy CID were not assignable using similar search parameters and therefore no clear results were obtainable. The knowledge of the properties of available MALDI-based fragmentation techniques presents an important factor for data interpretation using public accessible search algorithms and moreover for the identification of two-dimensional gel separated proteins.
机译：基于质谱的酶促生成肽测序被广泛用于获得特定序列标签，从而可以明确鉴定蛋白质。在本研究中，两种类型的解吸/电离技术结合不同的离子解离模式，即真空基质辅助激光解吸/电离（vMALDI）高能碰撞诱导解离（CID）和源后衰减（PSD）作为大气压（AP）-MALDI低能CID色谱柱，用于质子化的肽离子的片段化，这些离子来自二维分离的，银染的和胰蛋白酶消化的亲水性以及疏水性肾小球蛋白。因此，可以观察到由指定模式生成的各个碎片模式的定义属性。此外，已经评估了变化的PSD和CID（MS / MS）数据与使用两种公共可访问搜索算法的数据库搜索得出的标识的兼容性。在大多数情况下，通过PSD和高能CID获得的肽序列标签信息可实现明确识别。相反，通过低能量CID获得的部分数据无法使用相似的搜索参数进行分配，因此无法获得明确的结果。可用的基于MALDI的片段化技术特性的知识为使用公众可访问的搜索算法进行数据解释以及二维凝胶分离蛋白的鉴定提供了重要因素。

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