An 18O-labeling assisted LC/MS method for assignment of aspartyl/isoaspartyl products from Asn deamidation and Asp isomerization in proteins.
An 18O-labeling assisted LC/MS method for assignment of aspartyl/isoaspartyl products from Asn deamidation and Asp isomerization in proteins.
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DOI：
10.1021/ac400984r
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发表时间：
2013-07-02
影响因子：
7.4
通讯作者：
Kaltashov, Igor A.
中科院分区：
文献类型：
--
作者：
Wang, Shunhai;Kaltashov, Igor A.
设计了 O18 标记辅助 LC-MS 方法，用于明确分配 Asn 脱酰胺和 Asp 异构化产生的天冬氨酰/异天冬氨酰产物。通过在 H2O18 中制备酸和碱催化脱酰胺标准品，将异构体特异性质量标签引入到含天冬氨酰和异天冬氨酰的肽中，这些肽可以通过质谱 (MS) 轻松区分。与反相 HPLC 中根据洗脱顺序分配异构体的传统方法相比，新方法更加可靠和通用。此外，新方法可以应用于整个蛋白质消化，因此与使用合成的含天冬氨酰和异天冬氨酰的肽标准品的现有方法相比，更具时间和成本效益。最后，由于新方法中异构体的鉴定仅依赖于 LC-MS 分析，因此可以使用最基本且廉价的 MS 仪器轻松实现，从而为基于串联 MS 的方法提供了一种有吸引力的替代方案。使用模型肽以及人血清转铁蛋白的完整消化物证明了这种新方法的可行性。
An O18-labeling assisted LC-MS method was designed for unambiguous assignment of aspartyl/isoaspartyl products produced by Asn deamidation and Asp isomerization. By preparing the acid- and base–catalyzed deamidation standards in H2O18, isomer-specific mass tags were introduced to aspartyl- and isoaspartyl-containing peptides, which could be easily distinguished by mass spectrometry (MS). In contrast to the traditional ways to assign the isomers based on their elution order in reverse phase HPLC, the new method is more reliable and universal. Furthermore, the new method can be applied to the entire protein digest, and is therefore more time- and cost-effective compared with existing methods that use synthetic aspartyl- and isoaspartyl-containing peptide standards. Finally, since the identification of isomers in the new method only relies on LC-MS analysis, it can be easily implemented using the most basic and inexpensive MS instrumentation, thus providing an attractive alternative to tandem MS-based approaches. The feasibility of this new method is demonstrated using a model peptide as well as the entire digest of human serum transferrin.
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影响因子：
7.4
作者：
Gaza-Bulseco, Georgeen;Li, Biqin;Liu, Hongcheng
通讯作者：
Liu, Hongcheng
影响因子：
8
作者：
Cournoyer, JJ;Pittman, JL;O'Connor, PB
通讯作者：
O'Connor, PB
影响因子：
3
作者：
Kaltashov, Igor A.;Bobst, Cedric E.;Zhang, Mingxuan;Leverence, Rachael;Gumerov, Dmitry R.
通讯作者：
Gumerov, Dmitry R.
影响因子：
2.9
作者：
Noguchi, Shuji
通讯作者：
Noguchi, Shuji
DOI：
10.1073/pnas.1206924109
发表时间：
2012-08-21
影响因子：
11.1
作者：
Bobst, Cedric E.;Wang, Shunhai;Kaltashov, Igor A.
通讯作者：
Kaltashov, Igor A.