Exploring metallodrug-protein interactions by ESI mass spectrometry: The reaction of anticancer platinum drugs with horse heart cytochrome c
Exploring metallodrug-protein interactions by ESI mass spectrometry: The reaction of anticancer platinum drugs with horse heart cytochrome c
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DOI：
10.1002/cmdc.200500079
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发表时间：
2006-04-01
期刊：
影响因子：
3.4
通讯作者：
Pieraccini, Giuseppe
中科院分区：
文献类型：
--
作者：
Casini, Angela;Gabbiani, Chiara;Pieraccini, Giuseppe
由于DNA通常被认为是铂金属药物的主要靶点，[1-4]研究人员的兴趣主要集中在铂-核酸加合物的表征上，而对铂-蛋白质加合物的关注要少得多。然而，与蛋白质结合的铂片段可能代表真正有效的抗癌物种--而不仅仅是药物灭活产物--前提是不同结合部位之间的金属转移是动力学允许的。[4]此外，特定侧链的铂化，可以通过形成紧密的配位键影响生物关键蛋白质或酶的功能，可能在铂药物的整体机制和毒性中发挥相关作用。[5]铂-蛋白质相互作用的艺术状态在几篇文章和综述中描述；[6-8]无论如何，这一问题需要进一步的实验工作。由于最新的改进，今天的电喷雾电离质谱仪(ESI-MS)是探索金属药物-蛋白质相互作用的一种非常强大的方法。[6]由于引入了“软”电离方法，有可能在气相中转移完整的金属-蛋白质加合物--整个--以高精度测定其分子质量，从而获得其完整的分子特征。然而，针对这些系统的实验ESI-MS程序的优化和标准化仍然需要大量的工作。在目前的文献中，通常可以发现ESI-MS响应的很大的变异性，这取决于许多因素，例如蛋白质的性质、金属的性质、特定的溶液条件、金属结合配体的性质、pH和缓冲液的种类。显然，金属-蛋白质配位键的内在“脆弱性”是一个主要障碍，通常导致在电离过程中广泛的键断裂和化学信息的丢失。几年前，Gibson和他的同事报道了一些开创性的关于铂-蛋白质相互作用的ESI-MS研究，世界卫生组织使用泛素或肌红蛋白作为模型蛋白。[10]对一些铂蛋白加合物进行了详细的鉴定和表征。之后，其他研究小组又报道了几个不同金属药物-蛋白质加合物的ESI-MS研究。
Since DNA is commonly believed to be the primary target for platinum metallodrugs,[1–4] researchers’ interest has mainly focused on the characterisation of platinum–nucleic acid adducts while devoting much less attention to platinum–protein adducts. However, protein-bound platinum fragments probably represent truly active anticancer species—rather than mere drug-inactivation products—provided that metal transfer among distinct binding sites is kinetically allowed.[4] Moreover, platination of specific side chains, which can affect the function of biologically crucial proteins or enzymes through the formation of tight coordinative bonds, might play a relevant role in the overall mechanism and toxicity of platinum drugs.[5] The state of the art of platinum–protein interactions is described in a few articles and reviews;[6–8] in any case, this issue warrants further experimental work. Thanks to the latest improvements, electrospray ionisation mass spectrometry (ESI-MS) today represents a very powerful method for exploring metallodrug–protein interactions.[6] Owing to the introduction of “soft” ionisation methods, it is possible to transfer the intact metal–protein adduct—whole, in the gas phase—to determine its molecular mass with high accuracy and, thus, obtain its full molecular characterisation. However, much work is still required for the optimisation and the standardisation of experimental ESI-MS procedures directed at these systems. A great variability in ESI-MS responses is generally found in the current literature that depends on many factors, such as the nature of the protein, the nature of the metal, the specific solution conditions, the nature of the metalbound ligands, pH and the kind of buffer.[9] Apparently, the intrinsic “fragility” of the metal–protein coordination bonds represents a major obstacle, often leading to extensive bond cleavage during ionisation and to loss of chemical information.Some pioneering ESI-MS studies of platinum–protein interactions were reported a few years ago by Gibson and co-workers, who used either ubiquitin or myoglobin as model proteins.[10] A number of platinum–protein adducts were identified and characterised in detail. Afterwards, a few additional ESI-MS studies of various metallodrug–protein adducts were reported by other research groups.[11]