Identification and characterization of cysteinyl exposure in proteins by selective mercury labeling and nano-electrospray ionization quadrupole time-of-flight mass spectrometry
文献类型：期刊论文
| 作者 | Lu, Meiling ; Li, Xing-Fang ; Le, X. Chris ; Weinfeld, Michael ; Wang, Hailin |
| 刊名 | RAPID COMMUNICATIONS IN MASS SPECTROMETRY
 |
| 出版日期 | 2010-06 |
| 卷号 | 24期号:11页码:1523-1532 |
| 中文摘要 | We describe a method for probing surface-exposed cysteines in proteins by selective labeling with p-hydroxymercuribenzoate (FMB) combined with nano-electrospray ionization mass spectrometric analysis (nanoESI-MS). The rapid, stoichiometric, and specific labeling by PMB of surface-exposed cysteines allows for characterization of the accessibility of the cysteines using a single MS analysis. Moreover, by taking advantage of the large mass shift of 321 Da, unique isotopic pattern, and enhanced MS signal of PMB-labeled cysteine-containing peptide fragments, the surface-exposed cysteines in proteins can be accurately identified by peptide mapping. The number and sites of reactive cysteines on the surface of human and rat hemoglobins (hHb and rHb) were identified as examples. Collision-induced dissociation tandem mass spectrometric (MS/MS) analysis of specific peptides further confirmed the selective labeling of PMB in hHb. The subtle difference between the different cysteine residues in rHb was also evaluated by multiple PMB titrations. The difference between the two cysteines in their environment may partially explain their reaction specificity. Cysteine 1.25 in the beta unit of rHb is exposed on the surface, explaining its reactivity with glutathione. Cysteine 13 in the alpha subunit of rHb is much less exposed, and is located in a hydrophobic pocket, a conclusion that is consistent with the previous observation of its selective binding with dimethylarsinous acid, a reactive arsenic metabolite. The method is potentially useful for probing cysteines in other biologically important proteins and for studying proteins that are associated with conformational or structural changes induced by denaturing processes, protein modifications, protein-protein interactions and protein assemblies. Copyright (C) 2010 John Wiley & Sons, Ltd. |
| WOS记录号 | WOS:000278661200005 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/21087]  |
| 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,Li, Xing-Fang,Le, X. Chris,et al. Identification and characterization of cysteinyl exposure in proteins by selective mercury labeling and nano-electrospray ionization quadrupole time-of-flight mass spectrometry[J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY,2010,24(11):1523-1532. |
| APA | Lu, Meiling,Li, Xing-Fang,Le, X. Chris,Weinfeld, Michael,&Wang, Hailin.(2010).Identification and characterization of cysteinyl exposure in proteins by selective mercury labeling and nano-electrospray ionization quadrupole time-of-flight mass spectrometry.RAPID COMMUNICATIONS IN MASS SPECTROMETRY,24(11),1523-1532. |
| MLA | Lu, Meiling,et al."Identification and characterization of cysteinyl exposure in proteins by selective mercury labeling and nano-electrospray ionization quadrupole time-of-flight mass spectrometry".RAPID COMMUNICATIONS IN MASS SPECTROMETRY 24.11(2010):1523-1532. |
入库方式： OAI收割
来源：生态环境研究中心
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