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Study of metalation of thioredoxin by gold(I) therapeutic compounds using combined liquid chromatography/capillary electrophoresis with inductively coupled plasma/electrospray MS/MS detection

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Abstract

The reactivity of thioredoxin (Trx1) with the Au(I) drug auranofin (AF) and two therapeutic N-heterocyclic carbene (NHC)2-Au(I) complexes (bis [1-methyl-3-acridineimidazolin-2-ylidene]gold(I) tetrafluoroborate (Au3BC) and [1,3-diethyl-4,5-bis(4methoxyphenyl)imidazol-2-ylidene]gold(I) (Au4BC)) was investigated. Direct infusion (DI) electrospray ionization (ESI) mass spectrometry (MS) allowed information on the structure, stoichiometry, and kinetics of formation of Trx-Au adducts. The fragmentation of the formed adducts in the gas phase gave insights into the exact Au binding site within the protein, demonstrating the preference for Trx1 Cys32 or Cys35 of AF or the (NHC)2-Au(I) complex Au3BC, respectively. Reversed-phase HPLC suffered from the difficulty of elution of gold compounds, did not preserve the formed metal-protein adducts, and favored the loss of ligands (phosphine or NHC) from Au(I). These limitations were eliminated by capillary electrophoresis (CE) which enabled the separation of the gold compounds, Trx1, and the formed adducts. The ICP-MS/MS detection allowed the simultaneous quantitative monitoring of the gold and sulfur isotopes and the determination of the metallation extent of the protein. The hyphenation of the mentioned techniques was used for the analysis of Trx1-Au adducts for the first time.

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Funding

M. B. D. M. received from E2S-UPPA a PhD fellowship. Financial support of the CNR for “The Bioinorganic Drugs (BIDs) joint laboratory: A multidisciplinary platform promoting new molecular targets for drug discovery” is also received. M. M. and J. S. received from the Warsaw University of Technology financial support of the CE-ICP-MS/MS measurements.

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Authors and Affiliations

  1. Université de Pau Et Des Pays de L’Adour, E2S UPPA, CNRS, Institute of Analytical and Physical Chemistry for the Environment and Materials (IPREM-UMR 5254), 64053, Pau, France

    Mikel Bernabeu De Maria, Simon Godin, Joanna Szpunar, Ryszard Lobinski & Luisa Ronga

  2. Faculty of Chemistry, Warsaw University of Technology, Noakowskiego St. 3, 00-664, Warsaw, Poland

    Magdalena Matczuk, Jacek Sikorski & Ryszard Lobinski

  3. Department of Pharmacy and CIRPeB, Università degli Studi di Napoli Federico II, Via Montesano 49, 80131, Naples, Italy

    Diego Tesauro

  4. Instituto Di Cristallografia (IC), CNR, 70126, Caserta, Italy

    Michele Saviano

  5. Biological Mass Spectrometry and Proteomics (SMBP), ESPCI Paris, Université PSL, LPC CNRS UMR8249, 75005, Paris, France

    Giovanni Chiappetta

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  1. Mikel Bernabeu De Maria
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Correspondence to Luisa Ronga.

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Published in the topical collection Elemental Mass Spectrometry for Bioanalysis with guest editors Jörg Bettmer, Mario Corte-Rodríguez, and Márcia Foster Mesko.

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Bernabeu De Maria, M., Matczuk, M., Tesauro, D. et al. Study of metalation of thioredoxin by gold(I) therapeutic compounds using combined liquid chromatography/capillary electrophoresis with inductively coupled plasma/electrospray MS/MS detection. Anal Bioanal Chem 416, 2819–2833 (2024). https://doi.org/10.1007/s00216-024-05140-z

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