RT Journal Article
T1 Top-Down Mass Spectrometric Approach for the Full Characterization of Insulin-Cisplatin Adducts
A1 Moreno Gordaliza, María Estefanía
A1 Cañas Montalvo, Benito
A1 Gómez Gómez, María Milagros
A1 Gómez Gómez, M.Milagros
AB 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.
PB ACS
SN 1520-6882 (On line) 0003-2700 (Print)
YR 2009
FD 2009
LK https://hdl.handle.net/20.500.14352/52106
UL https://hdl.handle.net/20.500.14352/52106
LA eng
NO CICYT Projects
DS Docta Complutense
RD 8 abr 2025