Increasing the productivity of glycopeptides analysis by using higher-energy collision dissociation-accurate mass-product-dependent electron transfer dissociation.
Increasing the productivity of glycopeptides analysis by using higher-energy collision dissociation-accurate mass-product-dependent electron transfer dissociation.
复制标题
DOI：
10.1155/2012/560391
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Viner R
中科院分区：
文献类型：
--
作者：
Saba J;Dutta S;Hemenway E;Viner R
目前，聚糖作为潜在的生物标志物或作为治疗性蛋白质的翻译后修饰（PTM）引起了科学界的关注。然而，糖蛋白和糖肽的结构表征仍然具有分析挑战性。在这里，我们报告了一种新的采集策略，称为更高的能量碰撞解离精确的质量产品依赖的电子转移解离（HCD-PD-ETD）的混合线性离子阱轨道阱质谱仪的实施。该采集策略使用ETD和HCD的互补片段化以智能方式进行糖肽分析。此外，该方法最大限度地减少了优化仪器参数的用户输入，并能够直接检测糖肽。仅当检测到来自MS/MS HCD的聚糖氧鎓离子时才获得ETD光谱。这种方法的优点是它简化了数据分析，提高了动态范围和占空比。在这里，我们提出的好处HCD-PD-ETD相对于传统的交替HCD/ETD的教练组包含十二蛋白混合物与两种糖蛋白：人血清转铁蛋白，卵清蛋白和污染的其他两个：牛α 1酸性糖蛋白（bAGP）和牛胎球蛋白。
Currently, glycans are attracting attention from the scientific community as potential biomarkers or as posttranslational modifications (PTMs) of therapeutic proteins. However, structural characterization of glycoproteins and glycopeptides remains analytically challenging. Here, we report on the implementation of a novel acquisition strategy termed higher-energy collision dissociation-accurate mass-product-dependent electron transfer dissociation (HCD-PD-ETD) on a hybrid linear ion trap-orbitrap mass spectrometer. This acquisition strategy uses the complementary fragmentations of ETD and HCD for glycopeptides analysis in an intelligent fashion. Furthermore, the approach minimizes user input for optimizing instrumental parameters and enables straightforward detection of glycopeptides. ETD spectra are only acquired when glycan oxonium ions from MS/MS HCD are detected. The advantage of this approach is that it streamlines data analysis and improves dynamic range and duty cycle. Here, we present the benefits of HCD-PD-ETD relative to the traditional alternating HCD/ETD for a trainer set containing twelve-protein mixture with two glycoproteins: human serotransferrin, ovalbumin and contaminations of two other: bovine alpha 1 acid glycoprotein (bAGP) and bovine fetuin.