Identification of reactive cysteines in a protein using arsenic labeling and collision-induced dissociation tandem mass spectrometry
文献类型：期刊论文
| 作者 | Lu, Meiling ; Wang, Hailin ; Wang, Zhongwen ; Li, Xing-Fang ; Le, X. Chris |
| 刊名 | JOURNAL OF PROTEOME RESEARCH
 |
| 出版日期 | 2008-08 |
| 卷号 | 7期号:8页码:3080-3090 |
| 关键词 | hemoglobin arsenic mass defect labeling proteomics red blood cells protein binding site glutathione |
| 英文摘要 | Trivalent arsenicals have high affinity for thiols (such as free cysteines) in proteins. We describe here the use of this property to develop a collision-induced dissociation (CID) tandem mass spectrometry (MS/MS) technique for the identification of reactive cysteines in proteins. A trivalent arsenic species, dimethylarsinous acid (DMA(III)), with a residue mass (103.9607) and mass defect distinct from the normal 20 amino acids, was used to selectively label reactive cysteine residues in proteins. The CID fragment ions of the arsenic-labeled sequences shifted away from the more abundant normal fragments that would otherwise overlap with the ions of interest. Along with the internal and immonium ions, the arsenic-labeled fragment ions served as MS/MS signatures for identification of the binding sites and for assessment of the relative reactivity of individual cysteine residues in a protein. Using this method, we have identified two highly reactive binding sites in rat hemoglobin (Hb): Cys-13 alpha and Cys-125 beta. Cys-13 alpha was bound to DMA(III) in the Hb of rats fed with arsenic, and this binding was responsible for arsenic accumulation in rat blood, while Cys-125 beta was found to bind to glutathione in rat blood. This study revealed the relative reactivity of the cysteines in rat Hb in the following decreasing order: Cys-13 alpha >> Cys-111 alpha > Cys-104 alpha and Cys-13 alpha >> Cys-125 beta > Cys-93 beta. Arsenic-labeling is easy and fast for identification of active binding sites without enzymatic digestion and acid hydrolysis, and useful for characterization and identification of metal binding sites in other proteins. |
| WOS记录号 | WOS:000258200400003 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/21954]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Lu, Meiling,Wang, Hailin,Wang, Zhongwen,et al. Identification of reactive cysteines in a protein using arsenic labeling and collision-induced dissociation tandem mass spectrometry[J]. JOURNAL OF PROTEOME RESEARCH,2008,7(8):3080-3090. |
| APA | Lu, Meiling,Wang, Hailin,Wang, Zhongwen,Li, Xing-Fang,&Le, X. Chris.(2008).Identification of reactive cysteines in a protein using arsenic labeling and collision-induced dissociation tandem mass spectrometry.JOURNAL OF PROTEOME RESEARCH,7(8),3080-3090. |
| MLA | Lu, Meiling,et al."Identification of reactive cysteines in a protein using arsenic labeling and collision-induced dissociation tandem mass spectrometry".JOURNAL OF PROTEOME RESEARCH 7.8(2008):3080-3090. |
入库方式： OAI收割
来源：生态环境研究中心
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