ESI MS studies highlight the selective interaction of Auranofin with protein free thiols

Zoppi, Carlotta; Messori, Luigi; Pratesi, Alessandro

doi:10.1039/d0dt00283f

Cited by 61 publications

(72 citation statements)

References 55 publications

(69 reference statements)

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“…Thus, we performed ESI-MS experiments also for AF and AF-I incubated with the two latter amino acids. Results confirmed the lack of reactivity for these two target molecules with AF and AF-I, further supporting the findings already published by some of us regarding the reactivity of auranofin and its derivatives toward protein targets (Landini et al, 2017;Pratesi et al, 2018;Zoppi et al, 2020).…”

Section: Esi Mass Spectrometrysupporting

confidence: 90%

“…Starting from our previous findings on the reactivity of AF and its analogs toward protein targets (Pratesi et al, 2014(Pratesi et al, , 2018Marzo et al, 2017Marzo et al, , 2019Cirri et al, 2019;Zoppi et al, 2020), here we have extended the investigation-at the molecular levelon the mode of action of these medicinally relevant gold(I) compounds. Specifically, we have focused our attention on AF and its analog where the thiosugar moiety is replaced by the iodide ligand.…”

Section: Discussionmentioning

confidence: 99%

“…[Au(PEt 3 )] + is generated after release of the thiosugar ligand and it is capable to bind the key cellular targets triggering the pharmacological effect (Zou et al, 2000;Marzo et al, 2017Marzo et al, , 2019Zoppi et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

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Mechanistic Insights Into the Anticancer Properties of the Auranofin Analog Au(PEt3)I: A Theoretical and Experimental Study

et al. 2020

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Au(PEt 3)I (AF-I hereafter), the iodide analog of the FDA-approved drug auranofin (AF hereafter), is a promising anticancer agent that produces its pharmacological effects through interaction with non-genomic targets such as the thioredoxin reductase system. AF-I is endowed with a very favorable biochemical profile showing potent in vitro cytotoxic activity against several cancer types including ovarian and colorectal cancer. Remarkably, in a recent publication, some of us reported that AF-I induces an almost complete and rapid remission in an orthotopic in vivo mouse model of ovarian cancer. The cytotoxic potency does not bring about highly severe side effects, making AF-I very well-tolerated even for higher doses, even more so than the pharmacologically active ones. All these promising features led us to expand our studies on the mechanistic aspects underlying the antitumor activity of AF-I. We report here on an integrated experimental and theoretical study on the reactivity of AF-I, in comparison with auranofin, toward relevant aminoacidic residues or their molecular models. Results point out that the replacement of the thiosugar moiety with iodide significantly affects the overall reactivity toward the amino acid residues histidine, cysteine, methionine, and selenocysteine. Altogether, the obtained results contribute to shed light into the enhanced antitumoral activity of AF-I compared with AF.

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Section: Esi Mass Spectrometrysupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

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Mechanistic Insights Into the Anticancer Properties of the Auranofin Analog Au(PEt3)I: A Theoretical and Experimental Study

et al. 2020

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“…Briefly, the adopted experimental strategy was as follows. All four gold complexes were challenged with HSA, hCA I, and the TrxR dodecapeptide according to a well-established experimental protocol (Tamasi et al, 2014;Marzo et al, 2017Marzo et al, , 2019Michelucci et al, 2017;Pratesi et al, 2018;Ferraro et al, 2020;Zoppi et al, 2020). Different times and stoichiometries were explored to identify the best conditions to obtain adduct formation and to improve the quality of the ESI MS spectra.…”

Section: The Panel Of Gold(iii) Complexes and The Investigative Strategymentioning

confidence: 99%

“…This prompted investigators to analyze in detail the reactions of medicinal gold compounds with proteins. A systematic investigation on the interactions of the reference gold(I) drug auranofin with a series of model proteins, i.e., human serum albumin, carbonic anhydrase, superoxide dismutase, and hemoglobin, was recently carried out in our laboratory through mass spectrometry, and the resulting adducts could be characterized in molecular detail (Pratesi et al, 2016(Pratesi et al, , 2018(Pratesi et al, , 2019Magherini et al, 2018;Cirri et al, 2019;Zoppi et al, 2020). These studies revealed that auranofin targets quite selectively the free cysteine residues of proteins (Pratesi et al, 2018;Zoppi et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Reactions of Medicinal Gold(III) Compounds With Proteins and Peptides Explored by Electrospray Ionization Mass Spectrometry and Complementary Biophysical Methods

et al. 2020

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Electrospray ionization mass spectrometry (ESI MS) is a powerful investigative tool to analyze the reactions of metallodrugs with proteins and peptides and characterize the resulting adducts. Here, we have applied this type of approach to four experimental anticancer gold(III) compounds for which extensive biological and mechanistic data had previously been gathered, namely, Auoxo6, Au 2 phen, AuL12, and Aubipyc. These gold(III) compounds were reacted with two representative proteins, i.e., human serum albumin (HSA) and human carbonic anhydrase I (hCA I), and with the C-terminal dodecapeptide of thioredoxin reductase. ESI MS analysis allowed us to elucidate the nature of the resulting metal-protein adducts from which the main features of the occurring metallodrug-protein reactions can be inferred. In selected cases, MS data were integrated and supported by independent 1 HNMR and UV-Vis absorption measurements to gain an overall description of the occurring processes. From data analysis, it emerges that most of the investigated gold(III) complexes, endowed with an appreciable oxidizing character, undergo quite facile reduction to gold(I); the resulting gold(I) species tightly associate with the above proteins/peptides with a remarkable selectivity for free cysteine residues. In contrast, in the case of the less-oxidizing Aubipyc complex, the gold(III) oxidation state is conserved, and a gold(III) fragment still containing the original ligand is found to be associated with the target proteins. It is notable that the C-terminal dodecapeptide of thioredoxin reductase containing the characteristic-Gly-Cys-Sec-Gly metal-binding motif is able in all cases to trigger gold(III)-to-gold(I) reduction. Our investigation allowed us to identify in detail the nature of the gold fragments that ultimately bind the protein targets and determine the exact binding stoichiometry; some insight on the reaction kinetics was also gained. Notably, a few clear correlations could be established between the structure of the metal complexes and the nature of the resulting protein adducts. The mechanistic implications of these findings are analyzed and thoroughly discussed. Overall, the present results set the stage to better understand the real target biomolecules of these gold compounds and elucidate at the atomic level their interaction modes with proteins and peptides.

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Unlocking the Power of Human Ferritin: Enhanced Drug Delivery of Aurothiomalate in A2780 Ovarian Cancer Cells

Cosottini,

Geri,

Ghini

et al. 2024

Angewandte Chemie

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Aurothiomalate (AuTM) is an FDA‐approved antiarthritic gold drug with unique anticancer properties. To enhance its anticancer activity, we prepared a bioconjugate with human apoferritin (HuHf) by attaching some AuTM moieties to surface protein residues. The reaction of apoferritin with excess AuTM yielded a single adduct, that was characterized by ESI MS and ICP‐OES analysis, using three mutant ferritins and trypsinization experiments. The adduct contains ~3 gold atoms per ferritin subunit, arranged in a small cluster bound to Cys90 and Cys102. MD simulations provide a plausible structural model for the cluster. The adduct was evaluated for its pharmacological properties and was found to be significantly more cytotoxic than free AuTM against A2780 cancer cells mainly due to higher gold uptake. NMR‐metabolomics showed that AuTM bound to HuHf and free AuTM induced qualitatively similar changes in treated cancer cells, indicating that the effects on cell metabolism are approximately the same, in agreement with independent biochemical experiments. In conclusion, we have demonstrated here that a molecularly precise bioconjugate formed between AuTM and HuHf exhibits anticancer properties far superior to the free drug, while retaining its key mechanistic features. Evidence is provided that human ferritin can serve as an excellent carrier for this metallodrug.

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ESI MS studies highlight the selective interaction of Auranofin with protein free thiols

Abstract: The study of the mode-of-action of Auranofin, a cytotoxic gold(i) compound, reveals that it binds exclusively to the free and solvent-accessible cysteine residues of biologically relevant proteins.

Cited by 61 publications

References 55 publications

Mechanistic Insights Into the Anticancer Properties of the Auranofin Analog Au(PEt3)I: A Theoretical and Experimental Study

Mechanistic Insights Into the Anticancer Properties of the Auranofin Analog Au(PEt3)I: A Theoretical and Experimental Study

Reactions of Medicinal Gold(III) Compounds With Proteins and Peptides Explored by Electrospray Ionization Mass Spectrometry and Complementary Biophysical Methods

Unlocking the Power of Human Ferritin: Enhanced Drug Delivery of Aurothiomalate in A2780 Ovarian Cancer Cells

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