Systematic Identification of Proteins Binding with Cisplatin in Blood by Affinity Chromatography and a Four-Dimensional Proteomic Method

Abstract

Cisplatin is widely used for the treatment of various solid tumors. It is mainly administered by intravenous injection, and a substantial amount of the drug will bind to plasma proteins, a feature that is closely related to its pharmacokinetics, activity, toxicity, and side effects. However, due to the unique properties of platinum complexes and the complexity of the blood proteome, existing methods cannot systematically identify the binding proteome of cisplatin in blood. In this study, high-abundance protein separation and an ion mobility mass spectrometry-based 4D proteomic method were combined to systematically and comprehensively identify the binding proteins of cisplatin in blood. The characteristic isotope patterns of platinated peptides and a similarity algorithm were utilized to eliminate false-positive identification. Finally, 39 proteins were found to be platinated. Bioinformatics analysis showed that the identified proteins were mainly involved in the complement and coagulation cascade pathways. The binding ratio of some peptides with cisplatin was measured based on the area ratio of the free peptide using the parallel reaction monitoring method. This study provides a new method for systematically identifying binding proteins of metal drugs in blood, and the identified proteins might be helpful for understanding the toxicity of platinum anticancer drugs.

Comprehensive and systematic characterization of multi-functionalized cisplatin nano-conjugate: from the chemistry and proteomic biocompatibility to the animal model

“…As described in the recent report of Tao et al, where some of the proteins found in this work, are also described in the protein corona of these novel IONPs. As has been observed in this same work, there is a great difference between the protein corona in the different fluids studied [34]. In this sense, a difference has also been found between the activity observed in the in vitro studies of this and previous reports [16] and the in vivo studies proposed here.…”

Pt(ii) complex containing the 1R,2R enantiomer of trans-1,2-diamino-4-cyclohexene ligand effectively and selectively inhibits the viability of aggressive pancreatic adenocarcinoma cells and alters their lipid metabolism

“…Previously described platinum drug interactions with plasma proteins 18 led us to determine serum protein-mediated uptake of the investigated Pt( ii ) compounds into PSN1 pancreatic adenocarcinoma cells. For this purpose, we incubated first the test compounds for 24 h at 37 °C with complete fetal bovine serum (FBS).…”

Innovation in Cross-Linking Mass Spectrometry Workflows: Toward a Comprehensive, Flexible, and Customizable Data Analysis Platform

“…In addition, there is an algorithm for automatically assigning metallization sites of known proteins, called Apm2s. 62 This tool could identify fragment ions of cross-linked peptides and can compare the isotopic distribution of theoretical simulations with actual fragment ions, which is very helpful for the exclusion of false positive data. However, since it can only identify a single platinum-containing peptide, it cannot be well applied to the identification of platinumcontaining cross-linked peptides; Besides, only a single spectrum can be identified for a process, which takes a long time and cannot be batched, making it inefficient.…”

“…For these reasons, search engines designed for MS-cleavable cross-linkers such as XlinkX and MaxLynx, which rely on characteristic fragment ion peaks for peptide identification, are not compatible with platinum-based cross-linkers (Figure C). In addition, there is an algorithm for automatically assigning metallization sites of known proteins, called Apm2s . This tool could identify fragment ions of cross-linked peptides and can compare the isotopic distribution of theoretical simulations with actual fragment ions, which is very helpful for the exclusion of false positive data.…”