紫外激光诱导多肽的交联。
Ultraviolet laser-induced cross-linking in peptides.
机构信息
Dipartimento di Scienze Chimiche, Università di Napoli Federico II, Complesso Universitario di Monte S.Angelo, Napoli, Italy.
出版信息
Rapid Commun Mass Spectrom. 2013 Jul 30;27(14):1660-8. doi: 10.1002/rcm.6610.
RATIONALE
The aim of this study was to demonstrate, and to characterize by high-resolution mass spectrometry that it is possible to preferentially induce covalent cross-links in peptides by using high-energy femtosecond ultraviolet (UV) laser pulses. The cross-link is readily formed only when aromatic amino acids are present in the peptide sequence.
METHODS
Three peptides, xenopsin, angiotensin I, and interleukin, individually or in combination, were exposed to high-energy femtosecond UV laser pulses, either alone or in the presence of spin trapping molecules, the reaction products being characterized by high resolution mass spectrometry.
RESULTS
High-resolution mass spectrometry and spin trapping strategies showed that cross-linking occurs readily, proceeds via a radical mechanism, and is the highly dominant reaction, proceeding without causing significant photo-damage in the investigated range of experimental parameters.
CONCLUSIONS
High-energy femtosecond UV laser pulses can be used to induce covalent cross-links between aromatic amino acids in peptides, overcoming photo-oxidation processes, that predominate as the mean laser pulse intensity approaches illumination conditions achievable with conventional UV light sources.
原理
本研究的目的是通过使用高能飞秒紫外 (UV) 激光脉冲来证明并通过高分辨率质谱法来表征，可以优先诱导肽中的共价交联。只有当肽序列中存在芳香族氨基酸时，才容易形成交联。
方法
将三种肽，即 Xenopsin、血管紧张素 I 和白细胞介素，单独或组合使用，暴露于高能飞秒 UV 激光脉冲下，无论是单独使用还是在自旋捕获分子存在的情况下，通过高分辨率质谱法对反应产物进行表征。
结果
高分辨率质谱法和自旋捕获策略表明，交联很容易发生，通过自由基机制进行，并且是高度占主导地位的反应，在研究的实验参数范围内进行，而不会造成明显的光损伤。
结论
高能飞秒 UV 激光脉冲可用于诱导肽中芳香族氨基酸之间的共价交联，克服了光氧化过程，这些过程在平均激光脉冲强度接近使用传统 UV 光源可实现的照明条件时占主导地位。
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