School of Chemical Sciences, University of Auckland, Auckland, New Zealand.
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Electrophoresis. 2019 Sep;40(18-19):2329-2335. doi: 10.1002/elps.201900070. Epub 2019 May 21.
Cisplatin and its second and third generation analogues are widely used in the treatment of cancer. To study their reactions with proteins, we present a method based on SDS-PAGE separation and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) for platinum detection in the reaction between human serum albumin (HSA) and cisplatin. We developed matrix-matched standards of HSA/cisplatin mixtures and used them to quantify the amount of adducts formed at different HSA:cisplatin ratios. We noted that cisplatin incubation with HSA resulted in the formation of higher order HSA n-mers, depending on the amount of cisplatin added. This caused a depletion of the HSA dimer bands, while the majority of HSA was present as the monomer. Inducing the formation of such higher molecular weight species may have an impact on the mode of action of metallodrugs.
顺铂及其第二代和第三代类似物被广泛用于癌症的治疗。为了研究它们与蛋白质的反应,我们提出了一种基于 SDS-PAGE 分离和激光烧蚀-电感耦合等离子体质谱(LA-ICP-MS)的方法,用于检测人血清白蛋白(HSA)与顺铂反应中的铂。我们开发了 HSA/顺铂混合物的基质匹配标准品,并使用它们来定量不同 HSA:顺铂比例下形成的加合物的量。我们注意到,顺铂与 HSA 孵育会导致形成更高阶的 HSA n 聚体,具体取决于添加的顺铂量。这导致 HSA 二聚体带的耗尽,而大多数 HSA 以单体形式存在。诱导形成这种高分子量物质可能会影响金属药物的作用模式。