title: Mass spectrometric studies on the interaction of cisplatin and insulin.
abstract: The interaction of antitumor drug, cisplatin (cis-[PtCl2(NH3)2], CDDP) with insulin from porcine pancreas has been investigated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and high resolution hybrid ion trap/time-of-flight mass spectrometry (MALIDI-TOF/TOF-MS and ESI-IT/TOF MS). The MALDI-TOF/TOF-MS results demonstrated that the presence of cisplatin complex resulted in the reduction of the disulfide bond in porcine pancreas after the incubations of the two substances were performed in vitro. It indicated that the presence of cisplatin would destroy the native configuration of insulin, which may lead to the inactivation of insulin. High resolution mass values and the characteristic isotopic pattern of the platinated insulin ions allowed the analysis of platinated mono-, di- and triadducts of cisplatin and insulin in the incubations under different conditions. The laser-induced dissociation of the monoadduct obtained in MALDI source was carried out and one platinum was found to bind to insulin B chain was determined. The platinum binding sites were further identified to be the N terminus (B chain), cysteine 7 (B chain) and cysteine 19 (B chain) residues by electrospray ionization tandem mass spectrometry. The identification of the interaction between insulin and cisplatin broadens the horizon of the knowledge in the interaction of the proteins and metallodrugs.
authors.affiliation: Department of Chemical and Biological Engineering, Institute of Biological Engineering, Zhejiang University, Hangzhou, 310027, Zhejiang, China.|Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yue
authors.affiliation: Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China.
authors.forename: Xinchi
authors.initials: X
authors.lastname: Yin
authors.affiliation: Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Yin
authors.affiliation: Department of Chemistry, Zhejiang University, Hangzhou, 310027, Zhejiang, China. panyuanjiang@zju.edu.cn.
authors.forename: Yuanjiang
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Chemical and Biological Engineering, Institute of Biological Engineering, Zhejiang University, Hangzhou, 310027, Zhejiang, China. lryang@zju.edu.cn.
authors.forename: Lirong
authors.initials: L
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry*; D001665:Binding Sites; D002945:Cisplatin / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D007328:Insulin / Q000737:chemistry* / Q000302:isolation & purification; D010179:Pancreas / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine
chemical_list: D000970:Antineoplastic Agents; D004220:Disulfides; D007328:Insulin; D002945:Cisplatin
keywords: Binding sites; Cisplatin; Insulin from porcine pancreas; Insulin–cisplatin adduct; Mass spectrometry