Cell response of Escherichia coli to cisplatin‐induced stress

Abstract

Cisplatin is undoubtedly one of the most common and successful anticancer drugs worldwide. Though its DNA-based mechanism of action is well established, the contribution of the proteome to this process remains unclear. The possible impact of particular Escherichia coli proteins on the cytostatic activity of cisplatin was the subject of this study. Our main focus was not only the "bottom-up" identification of novel cisplatin protein targets through LC/LC-MS/MS analysis, but also a label-free quantification of their regulation profile by spectral-counting. The regulation of two proteins, aconitate hydratase 2 and 60 kDa chaperonin 1, could be linked to a platinated amino acid in the protein sequence, whereas in the cases of 30S ribosomal protein S1 and enolase, it could be shown that cisplatin fragments are coordinated to an essential site for the functionality of the protein. Nucleoside triphosphate pyrophosphohydrolase (MazG) regulates the programmed cell death and was found to be platinated on the protein surface, which probably correlates with the established mode of action. A possible new chapter in the understanding of cisplatin's mechanism of action and its severe side effects is opened, since evidence is provided that platinated proteins are not only involved in cellular stress response but also in energy metabolism through glycolysis and catabolic processes, in gene regulatory mechanisms and protein synthesis.

LA-ICP-MS and nHPLC-ESI-LTQ-FT-MS/MS for the analysis of cisplatin–protein complexes separated by two dimensional gel electrophoresis in biological samples

“…These latter proteins were demonstrated to be able to coordinate to Pt in previous cisplatin protein-binding studies in E. coli cells. 20,21 Moreover, a high amount of actin was found in spots 13-15, which presented a high Pt signal. Interestingly, cisplatin has been found to bind to G-actin and F-actin, inducing conformational changes and depolymerisation of F-actins and inhibition of G-actins polymerization at high drug concentrations, probably due to Pt binding to sulfur-containing residues.…”

Quantification of bindings of organometallic ruthenium complexes to GSTπ by mass spectrometry

“…With the advantages of high sensitivity, low sample consumption and chemical specificity, electrospray ionization mass spectrometry (ESI-MS) has become one of the most powerful tools for characterization of interactions between metallodrugs or drug candidates and proteins [1][2][3][4], in particular for elucidation of binding sites of metals in the proteins [5][6][7][8][9][10][11][12][13]. However, the uncertain alteration on ionization efficiency of proteins/peptides due to metallation and the lack of suitable internal standards make it a challenge to quantify the bindings of metal complexes to a specific site (residue) of the targeted proteins by ESI-MS, though inductive coupling plasma mass spectrometry (ICP-MS) in combination with various chromatography separation techniques has been widely used to determine the binding stoichiometry of metallodrugs to whole proteins [1,2,[14][15][16][17].…”

Isolate Specific Cold Response of Yersinia enterocolitica in Transcriptional, Proteomic, and Membrane Physiological Changes

“…Recently, advances in proteomics and bioinformatics technologies provide clear information on protein expression in response to cold and other stresses. High-throughput comparative proteomics with label-free quantification enabled the parsing of various potential mechanisms and regulatory networks of stress response in E. coli, B. subtilis, Pseudomonas putida, and Yersinia ruckeri (Delumeau et al, 2011;Stefanopoulou et al, 2011;Herbst et al, 2015;Kumar et al, 2016).…”