Enhancing electrospray ionization efficiency of peptides by derivatization
Enhancing electrospray ionization efficiency of peptides by derivatization
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DOI：
10.1021/ac0602266
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发表时间：
2006-06-15
影响因子：
7.4
通讯作者：
Regnier, Fred
中科院分区：
文献类型：
--
作者：
Mirzaei, Hamid;Regnier, Fred
随着电喷雾离子化质谱的出现，世界被赋予了一种新的方式来看待复杂的肽混合物。通过其特征肽鉴定蛋白质需要电离通过蛋白水解衍生自蛋白质的肽的代表性部分。不幸的是，基质效应禁止许多肽的电喷雾电离。本文介绍了一种新的标记试剂，同时增加了一个永久的正电荷的肽，并增加其疏水性，以提高其电离效率的发展。标记剂用N-羟基琥珀酰亚胺预活化，与伯胺反应形成肽键。在最引人注目的情况下，肽ADRDQYELLCLDNTRKPVDEYK的电离效率在衍生化后增加了500倍，而不是电离效率几乎不受影响的其他肽。肽的电离效率一般通过衍生化提高约10倍。小于500 Da的肽通过衍生化经历了电离效率的最大增加。发现天然肽的电离效率差更多是由于其固有的结构特性而不是电离发生的基质。
With the advent of electrospray ionization mass spectrometry, the world was given a new way to look at complex peptide mixtures. Identification of proteins via their signature peptides requires ionization of a representative portion of the peptides derived from proteins by proteolysis. Unfortunately, matrix effects prohibited electrospray ionization of many peptides. This paper describes the development of a new labeling reagent that simultaneously adds a permanent positive charge to peptides and increases their hydrophobicity to enhance their ionization efficiency. The labeling agent is preactivated with N-hydroxysuccinimide to react with primary amines to form a peptide bond. In the most dramatic case, ionization efficiency of the peptide ADRDQYELLCLDNTRKPVDEYK increased 500-fold after derivatization as opposed to other peptides where ionization efficiency was impacted little. Ionization efficiency of peptides was enhanced roughly 10-fold in general by derivatization. Peptides of less than 500 Da experienced the greatest increase in ionization efficiency by derivatization. Poor ionization efficiency of native peptides was found to be due more to their inherent structural properties than the matrix in which ionization occurs.