Dissecting ITC data of metal ions binding to ligands and proteins
Dissecting ITC data of metal ions binding to ligands and proteins
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DOI：
10.1016/j.bbagen.2015.08.018
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发表时间：
2016-05-01
影响因子：
3
通讯作者：
Grossoehme, Nicholas E.
中科院分区：
文献类型：
--
作者：
Johnson, Rachel A.;Manley, Olivia M.;Grossoehme, Nicholas E.
背景：ITC是一种强大的技术，可以可靠地评估大范围结合事件的热力学基础。当涉及到金属离子时，由于不可避免的溶液化学，包括金属形态和各种连锁平衡，在评估数据时出现了复杂性。审查范围：本文确定了这些问题，提供了避免常见错误的建议，并指导读者通过ITC数据的数学处理，以达到一组描述相同化学事件的热力学状态函数，理想情况下，与溶液条件无关。此外，生物金属传感研究中常用的金属发色团被提出作为一个鲁棒系统来确定未知溶液竞争。主要结论：金属离子在ITC实验中出现了一些并发症。这篇综述提出了避免这些陷阱的策略，并提出并实验验证了这些系统中存在的反卷积复杂平衡的数学方法。一般意义：本综述讨论了金属基ITC实验中广泛存在的并发症。它为这个领域的新科学家提供了一个起点，并阐明了将帮助有经验的研究人员排除实验故障的问题。这篇文章是Fadi boub - abdallah编辑的题为《生物科学中的微量热法-原理和应用》特刊的一部分。(C) 2015 Elsevier B.V.版权所有
Background: ITC is a powerful technique that can reliably assess the thermodynamic underpinnings of a wide range of binding events. When metal ions are involved, complications arise in evaluating the data due to unavoidable solution chemistry that includes metal speciation and a variety of linked equilibria.Scope of review: This paper identifies these concerns, provides recommendations to avoid common mistakes, and guides the reader through the mathematical treatment of ITC data to arrive at a set of thermodynamic state functions that describe identical chemical events and, ideally, are independent of solution conditions. Further, common metal chromophores used in biological metal sensing studies are proposed as a robust system to determine unknown solution competition.Major conclusions: Metal ions present several complications in ITC experiments. This review presents strategies to avoid these pitfalls and proposes and experimentally validates mathematical approaches to deconvolute complex equilibria that exist in these systems.General significance: This review discusses the wide range of complications that exists in metal-based ITC experiments. It provides a starting point for scientists new to this field and articulates concerns that will help experienced researchers troubleshoot experiments. This article is part of a Special Issue entitled Microcalorimetry in the BioSciences - Principles and Applications, edited by Fadi Bou-Abdallah. (C) 2015 Elsevier B.V. All rights reserved.