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学科主题: 药学
题名:
Solution-Phase Chelators for Suppressing Nonspecific Protein-Metal Interactions in Electrospray Mass Spectrometry
作者: Pan, Jingxi1,2,3; Xu, Kun4,5; Yang, Xiaoda4,5; Choy, Wing-Yiu1,2; Konermann, Lars1,2
刊名: ANALYTICAL CHEMISTRY
发表日期: 2009-06-15
DOI: 10.1021/ac900423x
卷: 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
所属项目编号: 07-0134 ; 20603003
项目资助者: 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
WOS记录号: WOS:000266969700043
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/63472
Appears in Collections:北京大学药学院_期刊论文

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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

Recommended Citation:
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.
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