Immobilized Pepsin Microreactor for Rapid Peptide Mapping with Nanoelectrospray Ionization Mass Spectrometry
Immobilized Pepsin Microreactor for Rapid Peptide Mapping with Nanoelectrospray Ionization Mass Spectrometry
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DOI：
10.1007/s13361-014-1015-8
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Wood, Troy D.
中科院分区：
文献类型：
--
作者：
Long, Ying;Wood, Troy D.
大多数用于蛋白质消化的酶促微反应器基于胰蛋白酶，但是具有疏水片段的蛋白质可能由于缺乏Arg和Lys残基而难以消化。基于胃蛋白酶的微反应器，其特异性低于胰蛋白酶，可以克服这一挑战。在这里，一个集成的固定化胃蛋白酶微反应器（IPMR）/纳米电喷雾发射器检查其潜在的肽映射。对于肌红蛋白，在具有更好的序列覆盖度的溶液消化的千分之一时间内获得相等的序列覆盖度。虽然细胞色素c的序列覆盖率在这段短时间内小于溶液，但产生了更多高电荷的消化肽，并且在低分辨率下未识别出一些峰，这表明需要高分辨率质谱法来充分利用集成的IPMR/纳米电喷雾装置。
Most enzymatic microreactors for protein digestion are based on trypsin, but proteins with hydrophobic segments may be difficult to digest because of the paucity of Arg and Lys residues. Microreactors based on pepsin, which is less specific than trypsin, can overcome this challenge. Here, an integrated immobilized pepsin microreactor (IPMR)/nanoelectrospray emitter is examined for its potential for peptide mapping. For myoglobin, equivalent sequence coverage is obtained in a thousandth the time of solution digestion with better sequence coverage. While sequence coverage of cytochrome c is lesser than solution in this short duration, more highly-charged peptic peptides are produced and a number of peaks are unidentified at low-resolution, suggesting that high-resolution mass spectrometry is needed to take full advantage of integrated IPMR/nanoelectrospray devices.