Higher Temperature Porous Graphitic Carbon Separations Differentially Impact Distinct Glycopeptide Classes.
Higher Temperature Porous Graphitic Carbon Separations Differentially Impact Distinct Glycopeptide Classes.
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DOI：
10.1021/jasms.2c00249
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发表时间：
2023-01-04
影响因子：
3.2
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Delafield, Daniel G.;Miles, Hannah N.;Ricke, William A.;Li, Lingjun
关键词：
基于质谱的发现糖蛋白组学高度依赖于色谱法的使用，该色谱法适用于分析物保留和分离。当与已建立的固定相如反相和亲水相互作用液相色谱相比时，利用多孔石墨碳（PGC）的报告详述了其许多优点。最近的努力已经详细说明了多孔石墨碳在高通量糖蛋白质组学中的实用性，主要是通过在较高柱温下增强的剖析深度和液相分辨率。然而，增加柱温已显示在糖肽鉴定中会产生干扰效应。在此，我们进一步阐明了这一趋势，描述了定性和半定量的影响，提高柱温对糖肽鉴定率，信号强度，分辨率和光谱计数线性响应。通过分析富集的牛和人糖肽，显示具有高甘露糖和唾液酸化聚糖的物种分别最显著地受益于高柱温和遭受高柱温。这些结果提供了关于如何适当利用多孔石墨碳分离进行糖肽鉴定的见解，同时随着温度的变化引起了对定量和半定量无标记比较的关注。RAW MS糖蛋白组学数据可通过ProteomeXchange获得，标识符为PXD 034354。
Mass spectrometry-based discovery glycoproteomics is highly dependent on the use of chromatography paradigms amenable to analyte retention and separation. When compared against established stationary phases such as reversed phase and hydrophilic interaction liquid chromatography, reports utilizing porous graphitic carbon (PGC) have detailed its numerous advantages. Recent efforts have detailed the utility in porous graphitic carbon in high throughput glycoproteomics, principally through enhanced profiling depth and liquid phase resolution at higher column temperatures. However, increasing column temperature has shown to impart disparaging effects in glycopeptide identification. Herein we further elucidate this trend, describing qualitative and semi-quantitative effects of increased column temperature on glycopeptide identification rates, signal intensity, resolution, and spectral count linear response. Through analysis of enriched bovine and human glycopeptides, species with high mannose and sialylated glycans were shown to most significantly benefit and suffer from high column temperatures, respectively. These results provide insight as to how porous graphitic carbon separations may be appropriately leveraged for glycopeptide identification while raising concerns over quantitative and semi-quantitative label free comparisons as temperature changes. RAW MS glycoproteomic data are available via ProteomeXchange with identifier PXD034354.
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影响因子：
4.1
作者：
Alpert, Andrew J.
通讯作者：
Alpert, Andrew J.
影响因子：
2.9
作者：
Abou-Abbass H;Abou-El-Hassan H;Bahmad H;Zibara K;Zebian A;Youssef R;Ismail J;Zhu R;Zhou S;Dong X;Nasser M;Bahmad M;Darwish H;Mechref Y;Kobeissy F
通讯作者：
Kobeissy F
影响因子：
4.3
作者：
Delafield, Daniel G.;Miles, Hannah N.;Liu, Yuan;Ricke, William A.;Li, Lingjun
通讯作者：
Li, Lingjun
影响因子：
15
作者：
ASHWELL, M;GUO, XM;SINNOTT, ML
通讯作者：
SINNOTT, ML
影响因子：
7.4
作者：
Escobar, Edwin E.;Wang, Shuaishuai;Goswami, Rupanjan;Lanzillotti, Michael B.;Li, Lei;McLellan, Jason S.;Brodbelt, Jennifer S.
通讯作者：
Brodbelt, Jennifer S.