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Solution-Phase Chelators for Suppressing Nonspecific Protein-Metal Interactions in Electrospray Mass Spectrometry
Pan, Jingxi1,2,3; Xu, Kun4,5; Yang, Xiaoda4,5; Choy, Wing-Yiu1,2; Konermann, Lars1,2
刊名ANALYTICAL CHEMISTRY
2009-06-15
DOI10.1021/ac900423x
81期:12页:5008-5015
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]GAS-PHASE ; LIGAND COMPLEXES ; NANOELECTROSPRAY IONIZATION ; NONCOVALENT COMPLEXES ; ALPHA-LACTALBUMIN ; PROTHYMOSIN-ALPHA ; DISSOCIATION-CONSTANTS ; BINDING SITES ; CARBOHYDRATE COMPLEXES ; BETA-LACTOGLOBULIN
英文摘要

Protein-metal complexes may be transferred from solution into the gas phase by electrospray ionization (ESI), such that they can be directly analyzed by mass spectrometry (MS). In principle, therefore, ESI-MS represents a simple and elegant approach for gaining insights into the binding stoichiometry and affinity of these assemblies. Unfortunately, the formation of nonspecific metal adducts during ESI can be a severe problem, often leading to binding levels that are dramatically higher than those in bulk solution. Focusing on several calcium binding proteins as test systems, this work explores the suitability of different salts to serve as metal source. Despite their widespread use in previous ESI-MS studies, calcium chloride and acetate induce extensive nonspecific adduction. In contrast, much lower levels of artifactual metal binding are observed in the presence of calcium tartrate. In the case of high and intermediate affinity proteins, the resulting ESI-MS data are in excellent agreement with the calcium binding behavior in bulk solution. The situation is more challenging when studying proteins with very low affinities, but in the presence of tartrate qualitative information on protein-metal interactions can still be obtained. The beneficial effects of tartrate also extend to zinc binding experiments. This work does not directly explore the mechanism by which tartrate suppresses nonspecific metalation. However, it seems likely that weak chelators such as tartrate sequester metal ions within rapidly shrinking droplets during the final stages of ESI, thereby reducing nonspecific metal adduction to protein carboxylates. The use of tartrate and possibly other weak chelators will greatly enhance the reliability of future ESI-MS studies on the interactions of proteins with divalent metal ions.

语种英语
WOS记录号WOS:000266969700043
项目编号07-0134 ; 20603003
资助机构Natural Sciences and Engineering Research Council of Canada (NSERC) ; Canada Foundation for Innovation (CFI) ; Province of Ontario ; Canada Research Chairs Program ; New Century Excellent Talents in University ; National Natural Science Foundation of China
引用统计
被引频次:30[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/63472
专题北京大学药学院
北京大学药学院_化学生物学系
作者单位1.Univ Western Ontario, Dept Chem, London, ON N6A 5B7, Canada
2.Univ Western Ontario, Dept Biochem, London, ON N6A 5B7, Canada
3.Dalian Univ Technol, State Key Lab Fine Chem, Dalian 116012, Peoples R China
4.Peking Univ, Sch Pharmaceut Sci, Beijing 100083, Peoples R China
5.Peking Univ, Natl Res Labs Nat & Biomimet Drugs, Beijing 100083, Peoples R China
推荐引用方式
GB/T 7714
Pan, Jingxi,Xu, Kun,Yang, Xiaoda,et al. Solution-Phase Chelators for Suppressing Nonspecific Protein-Metal Interactions in Electrospray Mass Spectrometry[J]. ANALYTICAL CHEMISTRY,2009,81(12):5008-5015.
APA Pan, Jingxi,Xu, Kun,Yang, Xiaoda,Choy, Wing-Yiu,&Konermann, Lars.(2009).Solution-Phase Chelators for Suppressing Nonspecific Protein-Metal Interactions in Electrospray Mass Spectrometry.ANALYTICAL CHEMISTRY,81(12),5008-5015.
MLA Pan, Jingxi,et al."Solution-Phase Chelators for Suppressing Nonspecific Protein-Metal Interactions in Electrospray Mass Spectrometry".ANALYTICAL CHEMISTRY 81.12(2009):5008-5015.
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