Zhang Ningbo, Liu Huan, Cui Meng, Du Yonggang, Liu Zhiqiang, Liu Shuying
Changchun Center of Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 5625 Renmin Street, Changchun, 130022, People's Republic of China.
University of the Chinese Academy of Sciences, Beijing, 100049, People's Republic of China.
J Biol Inorg Chem. 2015 Jan;20(1):1-10. doi: 10.1007/s00775-014-1202-x. Epub 2014 Oct 25.
Cisplatin has been widely used in the chemotherapy of a variety of tumors, and the interactions of cisplatin with proteins play very important roles in its side effects and drug resistance, as well as its pharmacokinetics and the biodistribution. Insulin-like growth factor-1 (IGF-1) was found to be associated with the drug resistance of cisplatin. Here, the interaction between cisplatin and IGF-1 was investigated using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. IGF-1-Pt(NH3)Cl was the main mono-adduct and the trans labilization was important to the reaction between IGF-1 and cisplatin, while another special mono-adduct IGF-1-Pt(NH3)Cl2 was observed. The rapid and sensitive top-down mass spectrometry-based approach in linear ion trap mass spectrometer has been developed to identify the binding sites of cisplatin in IGF-1 directly without tedious enzyme digestion. Three binding sites (Met59, Arg56 and Cys6) of cisplatin in IGF-1 were determined. The results not only provide a rapid and efficient way to identify the platinum binding sites in proteins, but also indicate that the binding of cisplatin could promote the fragmentation of IGF-1 and the rupture of disulfide bond.
顺铂已广泛应用于多种肿瘤的化疗，顺铂与蛋白质的相互作用在其副作用、耐药性以及药代动力学和生物分布中起着非常重要的作用。胰岛素样生长因子-1（IGF-1）被发现与顺铂的耐药性有关。在此，使用电喷雾电离傅里叶变换离子回旋共振质谱法研究了顺铂与IGF-1之间的相互作用。IGF-1-Pt(NH3)Cl是主要的单加合物，反式活化对IGF-1与顺铂之间的反应很重要，同时还观察到另一种特殊的单加合物IGF-1-Pt(NH3)Cl2。已开发出基于线性离子阱质谱仪的快速灵敏的自上而下质谱方法，可直接鉴定顺铂在IGF-1中的结合位点，而无需繁琐的酶切。确定了顺铂在IGF-1中的三个结合位点（Met59、Arg56和Cys6）。这些结果不仅提供了一种快速有效的方法来鉴定蛋白质中的铂结合位点，还表明顺铂的结合可以促进IGF-1的碎片化和二硫键的断裂。
