Study of Metalation of Thioredoxin by Gold(I) Therapeutic Compounds Using Combined Liquid Chromatography/capillary Electrophoresis with Inductively Coupled Plasma/electrospray MS/MS Detection

Overview

Journal Anal Bioanal Chem

Specialty Chemistry

Date 2024 Jan 20

PMID 38244050

Authors

Mikel Bernabeu de Maria

Magdalena Matczuk

Diego Tesauro

Michele Saviano

Jacek Sikorski

Giovanni Chiappetta

Simon Godin

Joanna Szpunar

Ryszard Lobinski

Luisa Ronga

Affiliations

Soon will be listed here.

Abstract

The reactivity of thioredoxin (Trx1) with the Au(I) drug auranofin (AF) and two therapeutic N-heterocyclic carbene (NHC)-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)-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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