Improved methods for the enrichment and analysis of glycated peptides.
Improved methods for the enrichment and analysis of glycated peptides.
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DOI：
10.1021/ac801704j
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发表时间：
2008-12-15
影响因子：
7.4
通讯作者：
Metz, Thomas O.
中科院分区：
文献类型：
--
作者：
Zhang, Qibin;Schepmoes, Athena A.;Brock, Jonathan W. C.;Wu, Si;Moore, Ronald J.;Purvine, Samuel O.;Baynes, John W.;Smith, Richard D.;Metz, Thomas O.
组织蛋白的非酶糖化在糖尿病并发症的发生中具有重要意义。在这里，我们报告了改进的方法，用于富集和分析糖化肽，分别使用硼酸盐亲和色谱和电子转移解离质谱。通过用50 mM乙酸铵在线洗涤柱结合的糖化肽，然后用100 mM乙酸洗脱来代替离线洗脱步骤，改善糖化肽的富集。通过在数据依赖性采集期间仅考虑较高电荷（≥3）的前体离子，改进了通过MS/MS进行的糖化肽分析，这增加了糖化肽鉴定的数量。类似地，当以增强的分辨率获得MS调查扫描时，电子转移后补充碰撞活化（ETcaD）的使用导致更多的糖化肽鉴定。以正常MS调查扫描速率获取ETD-MS/MS数据，结合1+和2+前体离子的排斥，相对于ETcaD或增强的MS调查扫描速率，增加了鉴定的糖化肽的数量。最后，对胰蛋白酶、Arg-C和Lys-C的评价表明，胰蛋白酶消化糖化蛋白与Lys-C消化相当，并且在通过LC-MS/MS鉴定的糖化肽和相应糖化蛋白的数量方面均优于Arg-C。
Nonenzymatic glycation of tissue proteins has important implications in the development of complications of diabetes mellitus. Herein we report improved methods for the enrichment and analysis of glycated peptides using boronate affinity chromatography and electron-transfer dissociation mass spectrometry, respectively. The enrichment of glycated peptides was improved by replacing an off-line desalting step with an online wash of column-bound glycated peptides using 50 mM ammonium acetate, followed by elution with 100 mM acetic acid. The analysis of glycated peptides by MS/MS was improved by considering only higher charged (≥3) precursor ions during data-dependent acquisition, which increased the number of glycated peptide identifications. Similarly, the use of supplemental collisional activation after electron transfer (ETcaD) resulted in more glycated peptide identifications when the MS survey scan was acquired with enhanced resolution. Acquiring ETD-MS/MS data at a normal MS survey scan rate, in conjunction with the rejection of both 1+ and 2+ precursor ions, increased the number of identified glycated peptides relative to ETcaD or the enhanced MS survey scan rate. Finally, an evaluation of trypsin, Arg-C, and Lys-C showed that tryptic digestion of glycated proteins was comparable to digestion with Lys-C and that both were better than Arg-C in terms of the number of glycated peptides and corresponding glycated proteins identified by LC–MS/MS.
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影响因子：
4.4
作者：
Brock, JWC;Hinton, DJS;Ames, JM
通讯作者：
Ames, JM
影响因子：
2.3
作者：
Frolov, Andrej;Hoffmann, Peter;Hoffmann, Ralf
通讯作者：
Hoffmann, Ralf
影响因子：
2.1
作者：
Takeuchi, Masayoshi;Kikuchi, Seiji;Yamagishi, Sho-ichi
通讯作者：
Yamagishi, Sho-ichi
影响因子：
4.4
作者：
Zhang, Qibin;Tang, Ning;Metz, Thomas O.
通讯作者：
Metz, Thomas O.
DOI：
10.1073/pnas.0607084104
发表时间：
2007-02-13
影响因子：
11.1
作者：
Chi, An;Huttenhower, Curtis;Hunt, Donald F.
通讯作者：
Hunt, Donald F.