期刊
PROTEOMICS
卷 9, 期 6, 页码 1617-1631出版社
WILEY
DOI: 10.1002/pmic.200800391
关键词
Chlorotyrosine; Hydroxytryptophan; Hydroxytyrosine; Oxidative stress; Precursor ion scanning
资金
- BBSRC [BBE0154841, BBE015727]
- BBSRC/EPSRC [BBC5115721]
- Conseil Regional d'Auvergne
- Nuffield Foundation
- BBSRC [BB/E015727/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E015727/1, BB/C511572/1] Funding Source: researchfish
Protein oxidation is thought to contribute to a number of inflammatory diseases, hence the development of sensitive and specific analytical techniques to detect oxidative PTMs (oxPTMs) in biological samples is highly desirable. Precursor ion scanning for fragment ions of oxidized amino acid residues was investigated as a label-free MS approach to mapping specific oxPTMs in a complex mixture of proteins. Using HOCl-oxidized lysozyme as a model system, it was found that the immonium ions of oxidized tyrosine and tryptophan formed in MS2 analysis could not be used as diagnostic ions, owing to the occurrence of isobaric fragment ions from unmodified peptides. Using a double quadrupole linear ion trap mass spectrometer, precursor ion scanning was combined with detection of MS3 fragment ions from the immonium ions and collisionally-activated decomposition peptide sequencing to achieve selectivity for the oxPTMs. For chlorotyrosine, the immonium ion at 170.1 m/z fragmented to yield diagnostic ions at 153.1, 134.1, and 125.1 m/z, and the hydroxytyrosine immonium ion at 152.1 m/z gave diagnostic ions at 135.1 and 107.1 m/z. Selective MS3 fragment ions were also identified for 2-hydroxytryptophan and 5-hydroxytryptophan. The method was used successfully to map these oxPTMs in a mixture of nine proteins that had been treated with HOCl, thereby demonstrating its potential for application to complex biological samples.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据