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首页> 外文期刊>Analytical chemistry >Coumarin Tags for Improved Analysis of Peptides by MALDI-TOF MS and MS/MS.1.Enhancement in MALDI MS Signal Intensities
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Coumarin Tags for Improved Analysis of Peptides by MALDI-TOF MS and MS/MS.1.Enhancement in MALDI MS Signal Intensities

机译：香豆素标签可通过MALDI-TOF MS和MS / MS改进肽段分析1.增强MALDI MS信号强度

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The goal of this study was the development of N-terminal tags to improve peptide identification using high-throughput MALDI-TOF MS and MS/MS.The proposed tags,commercially available fluorescent derivatives of cou-marin,can be advantageous for peptide analysis in both MS and MS/MS modes.This paper,part 1,will focus on the influence of derivatization on the intensities of MALDI-TOF MS signals of peptides.Labeling peptides with tags containing the coumarin core was found to enhance the intensities of peptide peaks (in some cases over 40-fold) in MALDI-TOF MS using CHCA and 2,5-DHAP matrixes.The signal enhancement was found to be peptide- and matrix-dependent,being the most pronounced for hydro-philic peptides.No correlation was found between the UV absorptivity of the tags at the excitation wavelengths typical for UV-MALDI and the magnitude of the signal enhancement.Interestingly,peptides labeled with Alexa Fluor 350,a coumarin derivative containing a sulfo group (i.e.,bearing strong negative charge),showed a 5-15-fold increase in intensity of MALDI MS signal in the positive ion mode,relative to the underivatized peptides,when 2,5-DHAP was used as the matrix.The Alexa Fluor 350 tag yielded a significantly higher signal relative to that for the CAF tag,likely due to the increased hydrophobicity of the coumarin structure.With 2,5-DHB,a decrease of MALDI MS signal was observed for all coumarin-labeled peptides,again relative to the unlabeled species.These findings support the hypothesis that derivatization with coumarin,a relatively hydrophobic structure,improves incorporation of hydrophilic peptides into hydrophobic MALDI matrixes,such as CHCA and 2,5-DHAP.
机译：这项研究的目的是开发N末端标签，以使用高通量MALDI-TOF MS和MS / MS改进肽的鉴定。拟议的标签，商业上可买到的香豆素荧光衍生物，可有利于肽段分析。本文的第1部分将重点探讨衍生化对肽的MALDI-TOF MS信号强度的影响。发现标记带有香豆素核心标签的肽可增强肽峰的强度使用CHCA和2,5-DHAP基质在MALDI-TOF MS中（在某些情况下超过40倍）。发现信号增强是肽和基质依赖性的，对于亲水性肽最为明显。发现在标签在典型的UV-MALDI激发波长下的UV吸收率和信号增强的幅度之间。有趣的是，用Alexa Fluor 350标记的肽是一种含有磺基的香豆素衍生物（即带有强负离子）电荷），表明当以2,5-DHAP为基质时，在阳离子模式下，相对于未衍生肽，MALDI MS信号强度增加了5-15倍.Alexa Fluor 350标签产生了显着相对于CAF标签，该信号具有更高的信号，这可能是由于香豆素结构的疏水性增加所致。对于2,5-DHB，所有香豆素标记的肽（相对于未标记物种）均观察到MALDI MS信号的降低。这些发现支持了以下假设：香豆素具有相对疏水的结构，可以改善亲水肽在疏水性MALDI基质（如CHCA和2,5-DHAP）中的掺入。

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