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在蛋白质学自下而上的实验中,具有Asn-Gly序列的脱amid化的染色学特性

Quinn Neale¹, Haley Neustaeter², Vic Spicer³

¹Department of Chemistry, University of Manitoba, 360 Parker Building, 144 Dysart Road, Winnipeg R3T 2N2, Canada; Manitoba Centre for Proteomics and Systems Biology, 799 JBRC, 715 McDermot Avenue, Winnipeg R3E 3P4, Canada.

Journal of chromatography. A

|November 16, 2024

概括

研究人员探索了脱化,专注于区分阿斯帕拉金 (Asn) 与其异构体酸 (Asp) 和异酸 (isoAsp). 染色学方法根据结构揭示了不同的化模式,有助于复杂样品中的脱化分析.

科学领域:

蛋白质组学是指蛋白质组学.
分析化学分析化学
生物化学生物化学

背景情况:

阿斯巴拉金 (Asn) 脱化为酸 (Asp) 和异酸 (isoAsp) 是一种常见的翻译后修饰.
区分这些同位素对于准确的蛋白质组分析至关重要,但质谱学在复杂的样本和高通量场景中面临着挑战.
染色学方法为异构体的分化提供了补充信息.

研究的目的:

在逆相分离中研究Asn及其除化产品 (Asp,isoAsp) 的保留特性.
为了建立质结构和脱化异构体的化顺序之间的相关性.
评估这些发现对高通量蛋白质组分析的有用性.

主要方法:

逆相高性能液态色谱 (RP-HPLC) 使用酸作为离子配对修饰剂.
分析含有不同初级和二级结构的三性的Asn-Gly.
使用分析尺度UV-HPLC确认化趋势,并在20个不同的分离设置中进行评估.

主要成果:

基于结构观察到Asn,isoAsp和Asp的三种不同的化模式:随机卷 (Asn < isoAsp < Asp),螺旋 (isoAsp < Asn < Asp) 和N终端Asn (Asn < Asp < isoAsp).
这些结构依赖的化趋势在复杂的蛋白质组样本中得到了验证.
对于各种与2D LC-MS/MS相关的分离条件来说,在除化后的保留转移被量化.

结论:

染色体保留行为为区分Asen除氧化产品提供了有价值的见解.
的初级和二级结构显著影响了Asn,Asp和isoAsp的化顺序.
已建立的化模式和量化的保留转移增强了染色学方法在高通量蛋白质组学中进行脱amidation分析的能力.

关键词:

脱化脱化脱化类保留预测的预测.后翻译修改后的修改.

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