Shotgun Analysis of Rough-Type Lipopolysaccharides Using Ultraviolet Photodissociation Mass Spectrometry.
Shotgun Analysis of Rough-Type Lipopolysaccharides Using Ultraviolet Photodissociation Mass Spectrometry.
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DOI：
10.1021/acs.analchem.5b04218
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发表时间：
2016-01-05
影响因子：
7.4
通讯作者：
Brodbelt JS
中科院分区：
文献类型：
--
作者：
Klein DR;Holden DD;Brodbelt JS
完整的粗糙型脂多糖（R-LPS）的详细结构表征完成使用MS 3策略，包括碰撞诱导解离（CID），然后由193紫外光解离（UVPD）上实施的轨道阱融合质谱仪。来自E. coli或S.肠毒素直接注入质谱仪使用静态源nanoESI。在R-LPS前体上进行的初始CID事件产生具有对应于脂质A和核心寡糖（OS）亚结构的丰富离子的光谱。具有不同脂质A和核心OS结构的R-LPS离子的CID光谱的比较证实脂质A和核心OS离子以高丰度一致地产生。随后通过CID、高能碰撞诱导解离（HCD）或UVPD活化所得脂质A和核心OS离子。对于脂质A和核心OS亚结构，HCD和UVPD产生高度信息互补的光谱，与UVPD的核心OS产生广泛的交叉环切割片段阵列。成功识别E.具有异构核心结构的大肠杆菌R-LPS结构证实了使用该方法可以确定细微结构差异的程度。
Detailed structural characterization of intact rough-type lipopolysaccharides (R-LPS) was accomplished using an MS3 strategy consisting of collision-induced dissociation (CID) followed by 193 ultraviolet photodissociation (UVPD) implemented on an Orbitrap Fusion mass spectrometer. Complex mixtures of R-LPS from either E. coli or S. enterica were directly infused into the mass spectrometer using static source nanoESI. An initial CID event performed on an R-LPS precursor produced spectra with abundant ions corresponding to the lipid A and core oligosaccharide (OS) substructures. Comparison of CID spectra of R-LPS ions with varying lipid A and core OS structures verify that lipid A and core OS ions are consistently produced in high abundance. The resulting lipid A and core OS ions were subsequently activated by CID, high-energy collision induced dissociation (HCD), or UVPD. For both the lipid A and core OS substructures, HCD and UVPD produced highly informative complementary spectra, with UVPD of the core OS producing an extensive array of cross-ring cleavage fragments. Successful discernment of E. coli R-LPS structures with isomeric core structures confirmed the degree to which subtle structural differences could be determined using this method.