2026-06-27T16:26:09Zhttps://docta.ucm.es/rest/oai/request

oai:docta.ucm.es:20.500.14352/521062024-10-01T17:57:55Zcom_20.500.14352_14col_20.500.14352_15

Moreno Gordaliza, María Estefanía Cañas Montalvo, Benito Gómez Gómez, María Milagros Gómez Gómez, M.Milagros 2023-06-20T12:39:40Z 2023-06-20T12:39:40Z 2009 1520-6882 (On line) 0003-2700 (Print) 10.1021/ac900046v https://hdl.handle.net/20.500.14352/52106 http://pubs.acs.org/doi/abs/10.1021/ac900046v The interaction of the antitumor drug cisplatin with insulin was studied using a top-down mass spectrometric approach. In vitro incubations were prepared under acidic and physiological conditions at different insulin/cisplatin molar ratios for different incubation times. Size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICPMS) analysis enabled the specific detection of platinum containing species attributed to the binding of the drug to the protein. Further analysis through matrix-assisted laser desorption ionization-timeof-flight mass spectrometry (MALDI-TOF MS) and nanoelectrospray ionization mass spectrometry using a linear ion trap (nESI-LIT-MS) allowed the identification of platinated mono-, di-, and even triadducts in the incubations. Platinum binding sites were identified by CID-MSn as B chain N-terminus, His5, and probably His10 residues, which turned out to be the same, regardless of the incubation conditions. Evidence on the binding of Pt to B chain Cys7 was also observed. Working with the LIT zoom scan mode provides enough resolution to discern the isotopic pattern for both precursor and fragment ions, allowing the differentiation of platinumcontaining ions. The elucidation of platinum binding sites in a native protein through a top-down approach has been performed for the first time with this type of instrument. eng restricted access Top-Down Mass Spectrometric Approach for the Full Characterization of Insulin-Cisplatin Adducts journal article