Abstract:
Glycosylation is a common modification across living organisms and plays a central role in understanding biological systems and disease. Our ability to probe the gylcome has grown exponentially in the past several decades. However, further improvements to the analytical toolbox available to researchers would allow for increased capabilities to probe structure and function of biological systems and to improve disease treatment. This article applies the developing technique of two-dimensional Fourier transform ion cyclotron resonance mass spectrometry to a glycoproteomic workflow for the standard glycoproteins coral tree lectin (CTL) and bovine ribonuclease B (BRB) to demonstrate its feasibility as a tool for glycoproteomic workflows. 2D infrared multiphoton dissociation and electron capture dissociation spectra of CTL reveal comparable structural information to their 1D counterparts, confirming the site of glycosylation and monosaccharide composition of the glycan. Spectra collected in 2D of BRB reveal correlation lines of fragment ion scans and vertical precursor ion scans for data collected using infrared multiphoton dissociation and diagonal cleavage lines for data collected by electron capture dissociation. The use of similar techniques for glycoproteomic analysis may prove valuable in instances where chromatographic separation is undesirable or quadrupole isolation is insufficient.
摘要:
糖基化是生物体中常见的修饰,在理解生物系统和疾病中起着核心作用。在过去的几十年中,我们探测gylcome的能力呈指数级增长。然而,对研究人员可用的分析工具箱的进一步改进将允许增强探测生物系统的结构和功能以及改善疾病治疗的能力。本文将二维傅立叶变换离子回旋共振质谱技术应用于标准糖蛋白珊瑚树凝集素(CTL)和牛核糖核酸酶B(BRB)的糖蛋白质组学工作流程,以证明其作为糖蛋白质组学工作流程工具的可行性。CTL的2D红外多光子解离和电子捕获解离光谱揭示了与其1D对应物相当的结构信息,确认聚糖的糖基化位点和单糖组成。在BRB的2D中收集的光谱揭示了使用红外多光子解离收集的数据的碎片离子扫描和垂直前体离子扫描的相关线,以及通过电子捕获解离收集的数据的对角分裂线。在不希望色谱分离或四极杆分离不充分的情况下,使用类似的技术进行糖蛋白质组分析可能证明是有价值的。