Modelling the electrophoretic migration behaviour of peptides and glycopeptides from glycoprotein digests in capillary electrophoresis-mass spectrometry
Modelling the electrophoretic migration behaviour of peptides and glycopeptides from glycoprotein digests in capillary electrophoresis-mass spectrometry
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DOI：
10.1016/j.aca.2014.10.038
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发表时间：
2015-01-07
影响因子：
6.2
通讯作者：
Sanz-Nebot, Victoria
中科院分区：
文献类型：
--
作者：
Barroso, Albert;Gimenez, Estela;Sanz-Nebot, Victoria
在这项研究中，利用电泳迁移率和荷质比之间的经典半经验关系（$μ_e$对$q/M^{-\alpha}$）来模拟源自重组人促红细胞生成素（rhEPO，一种具有生物学和治疗相关性的糖蛋白）消化产物的肽段和糖肽的迁移行为。对斯托克斯定律（$\alpha = 1/3$）、经典聚合物模型（$\alpha = 1/2$）和奥福德表面定律（$\alpha = 2/3$）进行评估，以预测用胰蛋白酶和胰蛋白酶 - 神经氨酸酶消化的rhEPO中含或不含唾液酸（SiA）的肽段和糖肽的迁移情况。对于用于评估模型的一组肽段，斯托克斯定律得出了更好的相关性，而糖肽则更符合经典聚合物模型。一旦用这两个模型预测了迁移时间，就很容易模拟它们的分离电泳图。随后通过预测不同组的rhEPO糖肽以及人转铁蛋白（Tf）肽段和糖肽的迁移并模拟其分离来验证结果。rhEPO和Tf消化产物的实验电泳图和模拟电泳图之间的高度一致性证实了这种简单策略在一般情况下预测任何消化后的糖蛋白的肽 - 糖肽电泳图谱的潜在适用性。（C）2014年由爱思唯尔出版集团出版。
In this study, the classical semiempirical relationships between the electrophoretic mobility and the charge-to-mass ratio (me vs. q/M-alpha) were used to model the migration behaviour of peptides and glycopeptides originated from the digestion of recombinant human erythropoietin (rhEPO), a biologically and therapeutically relevant glycoprotein. The Stoke's law (alpha = 1/3), the classical polymer model (alpha = 1/2) and the Offord's surface law (alpha = 2/3) were evaluated to predict migration of peptides and glycopeptides, with and without sialic acids (SiA), in rhEPO digested with trypsin and trypsin-neuraminidase. The Stoke's law resulted in better correlations for the set of peptides used to evaluate the models, while glycopeptides fitted better with the classical polymer model. Once predicted migration times with both models, it was easy to simulate their separation electropherogram. Results were later validated predicting migration and simulating separation of a different set of rhEPO glycopeptides and also human transferrin (Tf) peptides and glycopeptides. The excellent agreement between the experimental and the simulated electropherograms with rhEPO and Tf digests confirmed the potential applicability of this simple strategy to predict, in general, the peptide-glycopeptide electrophoretic map of any digested glycoprotein. (C) 2014 Published by Elsevier B.V.