Ultrafast ultraviolet photodissociation at 193 nm and its applicability to proteomic workflows.
Ultrafast ultraviolet photodissociation at 193 nm and its applicability to proteomic workflows.
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DOI：
10.1021/pr100515x
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发表时间：
2010-08-06
影响因子：
4.4
通讯作者：
Brodbelt JS
中科院分区：
文献类型：
--
作者：
Madsen JA;Boutz DR;Brodbelt JS
在一台线性离子阱质谱仪上实施了193纳米的紫外光解离（UVPD），用于高通量蛋白质组学工作流程。在单个5纳秒激光脉冲照射下，实现了胰蛋白酶肽的高效光解离，除了产生铵离子以及v和w侧链丢失离子外，还产生了a、b、c、x、y和z系列离子。对影响UVPD质谱以及随后通过SEQUEST进行计算机数据库检索的因素进行了评估。发现肽序列的芳香性和前体电荷态对光解离效率的影响比酰胺生色团的数量更大，并且离子阱q值和激光脉冲数量显著影响诊断产物离子（例如系列离子和铵离子）的数量和丰度。此外，光致电离背景扣除被证明可极大地改善SEQUEST结果，尤其是当肽信号较低时。实施了一种液相色谱 - 质谱（LC - MS） - UVPD策略，并且在分析蛋白水解的牛血清白蛋白和裂解的人HT - 1080细胞溶质纤维肉瘤细胞时，相对于LC - MS - 碰撞诱导解离（CID）产生了相当或更好的结果。
Ultraviolet photodissociation (UVPD) at 193 nm was implemented on a linear ion trap mass spectrometer for high-throughput proteomic workflows. Upon irradiation by a single 5 ns laser pulse, efficient photodissociation of tryptic peptides was achieved with production of a, b, c, x, y, and z sequence ions, in addition to immonium ions and v and w side-chain loss ions. The factors that influence the UVPD mass spectra and subsequent in silico database searching via SEQUEST were evaluated. Peptide sequence aromaticity and the precursor charge state were found to influence photodissociation efficiency more so than the number of amide chromophores, and the ion trap q-value and number of laser pulses significantly affected the number and abundances of diagnostic product ions (e.g., sequence and immonium ions). Also, photoionization background subtraction was shown to dramatically improve SEQUEST results, especially when peptide signals were low. A liquid chromatography – mass spectrometry (LC-MS) – UVPD strategy was implemented and yielded comparable or better results relative to LC-MS – collision induced dissociation (CID) for analysis of proteolyzed bovine serum albumin and lysed human HT-1080 cytosolic fibrosarcoma cells.
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影响因子：
2.9
作者：
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通讯作者：
Glish, Gary L.
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Zabrouskov, Vlad
DOI：
10.1016/j.jasms.2009.08.021
发表时间：
2009-12-01
影响因子：
3.2
作者：
Kim, Tae-Young;Reilly, James P.
通讯作者：
Reilly, James P.