Targeting Cysteine Residues of Biomolecules: New Approaches for the Design of Antiviral and Anticancer Drugs

Scozzafava, A.; Casini, Angela; Supuran, Claudiu T.

doi:10.2174/0929867023370077

Cited by 52 publications

(34 citation statements)

References 80 publications

(228 reference statements)

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“…Thus, selectivity of adduction is governed by both the recipient nucleophile and the chemistry of the adducting electrophile. Likewise, Cys-239 in tubulin is preferentially alkylated by anticancer drugs, including the colchicines, Vinca alkaloids, rhizoxin/maytansine, and taxanes (Jordan et al, 1998;Shan et al, 1999;Kavallaris et al, 2001;Casini et al, 2002;Scozzafava et al, 2002), and carbonic anhydrase III, which contains five cysteine residues, is selectively alkylated at Cys-186 by acrylonitrile (Nerland et al, 2003). Cysteine thiols are clearly common targets of reactive electrophilic metabolites and endogenous electrophiles, including lipid-derived ␣,␤-unsaturated aldehydes, such as 4-hydroxynonenal (4-HNE) and 4-oxononenal, and reactive oxygen species.…”

Section: Discussionmentioning

confidence: 99%

Protein Electrophile-Binding Motifs: Lysine-Rich Proteins Are Preferential Targets of Quinones

Labenski¹,

Fisher²,

Lo³

et al. 2009

Drug Metab Dispos

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ABSTRACT:Quinones represent an important class of endogenous compounds such as neurotransmitters and coenzyme Q10, electrophilic xenobiotics, and environmental toxicants that have known reactivity based on their ability to redox cycle and generate oxidative stress, as well as to alkylate target proteins. It is likely that topological, chemical, and physical features combine to determine which proteins become targets for chemical adduction. Chemical-induced post-translational modification of certain critical proteins causes a change in structure/function that contributes to the toxicological response to chemical exposure. In this study, we have identified a number of proteins that are modified by quinone-thioethers after administration of 2-(glutathion-S-yl)HQ. Parallel one-dimensional gel electrophoresis was performed, and the Coomassie-stained gel was aligned with the corresponding Western blot, which was probed for adductions. Immunopositive bands were then subjected to trypsin digestion and analyzed via liquid chromatography/tandem mass spectrometry. The proteins that were subsequently identified contained a higher than average (9.7 versus 5.5%) lysine content and numerous stretches of lysine run-ons, which is a presumed electrophile binding motif. Approximately 50% of these proteins have also been identified as targets for electrophilic adduction by a diverse group of chemicals by other investigators, implying overlapping electrophile adductomes. By identifying a motif targeted by electrophiles it becomes possible to make predictions of proteins that may be targeted for adduction and possible sites on these proteins that are adducted. An understanding of proteins targeted for adduction is essential to unraveling the toxicity produced by these electrophiles.Proteins have long been recognized as being critical targets of chemicals that produce acute tissue toxicities and cancer. Moreover, the adverse effects of many drugs are frequently mediated via their metabolic activation to reactive electrophilic metabolites, which subsequently bind covalently to proteins. Such reactive metabolites usually have low electron density and react with molecular centers of high electron density (i.e., nucleophiles). Target proteins for adduction usually contain strong nucleophilic sites, including cysteine thiols, lysine amines, histidine imidazoles, and protein N-terminal amines, which are readily attacked by reactive species (Guengerich et al., 2001;Casini et al., 2002). Other proteins contain weaker nucleophilic sites, including methionine sulfur, arginine guanidinium, tyrosine phenols, serine and threonine hydroxyls, and aspartate and glutamate carboxyls.Attempts to identify protein targets of reactive electrophilic metabolites have been complicated because 1) proteins have tremendously diverse structures and properties, 2) unmodified proteins are frequently present in excess, and 3) radiolabeled chemicals of sufficiently high specific activity are often unavailable or prohibitively expensive. Antibody-based approaches to a...

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Section: Discussionmentioning

confidence: 99%

Protein Electrophile-Binding Motifs: Lysine-Rich Proteins Are Preferential Targets of Quinones

Labenski¹,

Fisher²,

Lo³

et al. 2009

Drug Metab Dispos

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“…Thus, it may be argued that disulfiram is akin to the failed first-generation modulators of Pgp, which were originally developed for other disease conditions and for reversing MDR only at relatively high concentrations. However, the extremely low toxicity of disulfiram (Chick, 1999) coupled with the fact that sulfhydryl-reacting agents, despite their apparently unspecific mode of action, provide useful drugs against human diseases (Yakisich et al, 2001;Scozzafava et al, 2002) distinguishes it from other first-generation MDR modulators. Also, disulfiram has been in clinical use for several decades and has a well-documented pharmacology.…”

Section: Discussionmentioning

confidence: 99%

The Molecular Basis of the Action of Disulfiram as a Modulator of the Multidrug Resistance-Linked ATP Binding Cassette Transporters MDR1 (ABCB1) and MRP1 (ABCC1)

Sauna¹,

Peng²,

Nandigama³

et al. 2004

Mol Pharmacol

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The overexpression of multidrug resistance protein 1 (MDR1) and multidrug resistance protein 1 (MRP1) gene products is a major cause of multidrug resistance in cancer cells. A recent study suggested that disulfiram, a drug used to treat alcoholism, might act as a modulator of P-glycoprotein. In this study, we investigated the molecular and chemical basis of disulfiram as a multidrug resistance modulator. We demonstrate that in intact cells, disulfiram reverses either MDR1-or MRP1-mediated efflux of fluorescent drug substrates. Disulfiram inhibits ATP hydrolysis and the binding of [␣-32 P]8-azidoATP to Pglycoprotein and MRP1, with inhibition curves comparable with those of N-ethylmaleimide, a cysteine-modifying agent. However, if the ATP sites are protected with excess ATP, disulfiram stimulates ATP hydrolysis by both transporters in a concentration-dependent manner. Thus, in addition to modifying cysteines at the ATP sites, disulfiram may interact with the drugsubstrate binding site. We demonstrate that disulfiram, but not N-ethylmaleimide, inhibits in a concentration-dependent manner the photoaffinity labeling of the multidrug transporter with 125 I-iodoarylazidoprazosin and [3 H]azidopine. This suggests that the interaction of disulfiram with the drug-binding site is independent of its role as a cysteine-modifying agent. Finally, we have exploited MRP4 (ABCC4) to demonstrate that disulfiram can inhibit ATP binding by forming disulfide bonds between cysteines located in the vicinity of, although not in, the active site. Taken together, our results suggest that disulfiram has unique molecular interactions with both the ATP and/or drug-substrate binding sites of multiple ATP binding cassette transporters, which are associated with drug resistance, and it is potentially an attractive agent to combat multidrug resistance.

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“…In addition to activating intracellular oxidant stress pathways, mass spectrometry (MS) analysis has recently revealed that brief exposure to K3 can result in arylation of the singular cysteine residue (Cys110) within histone H3.3/H3 [16]. Earlier studies had shown that brief treatment of ER-expressing breast cancer cells in culture with K3 also results in irreversible functional loss of ER-DNA binding [3], presumably due to the Michael addition of either the quinone or hydroquinone form of K3 to one or more nucleophilic groups within the ER-DBD [17,18]. However, analytical and structural studies of chemical modifications within the ER-DBD remain challenging, given the much lower abundance of endogenous ER relative to proteins like histone H3.3/H3, even when ER is markedly overexpressed in human breast cancer cells.…”

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confidence: 99%

Reactivity of zinc finger cysteines: Chemical modifications within labile zinc fingers in estrogen receptor

Atsriku

Scott

Benz

et al. 2005

J. Am. Soc. Mass Spectrom.

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Estrogen receptor (ER, alpha isoform) is a 67 kDa zinc finger transcription factor that plays a fundamental role in both normal reproductive gland development and breast carcinogenesis, and also represents a critical molecular target for breast cancer therapy. We are investigating the structural consequences of chemical exposures thought to modify essential zinc finger cysteine residues in human ER. The current study employs mass spectrometry to probe ER zinc finger structural changes induced by a redox-reactive vitamin K3 analog, menadione; a commonly used cysteine alkylator, iodoacetic acid; and a thiol alkylating fluorophore, monobromobimane. Although they are slower to react, the sterically bulkier reagents, monobromobimane and menadione, effectively alkylate the most susceptible ER zinc finger cysteine sulfhydryl groups. Menadione arylation results first in Michael addition of the hydroquinone followed by rapid oxidation to the corresponding quinone, evidenced by a 2 Da mass loss per cysteine residue. Mass spectrometric analysis performed under MALDI conditions reveals both hydroquinone and quinone forms of arylated menadione, whereas only the quinone product is detectable under ESI conditions. Tandem mass spectrometry of a synthetic peptide encompassing the C-terminal half of the structurally more labile second zinc finger of ER (ZnF2B) demonstrates that the two nucleophilic thiols in ZnF2B (Cys-237, Cys-240) are not chemically equivalent in their reactivity to bromobimane or menadione, consistent with their unequal positioning near basic amino acids that affect thiol pKa, thereby rendering Cys-240 more reactive than Cys-237. These findings demonstrate important differential susceptibility of ER zinc finger cysteine residues to thiol reactions. (J Am Soc Mass Spectrom

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Targeting Cysteine Residues of Biomolecules: New Approaches for the Design of Antiviral and Anticancer Drugs

Cited by 52 publications

References 80 publications

Protein Electrophile-Binding Motifs: Lysine-Rich Proteins Are Preferential Targets of Quinones

Protein Electrophile-Binding Motifs: Lysine-Rich Proteins Are Preferential Targets of Quinones

The Molecular Basis of the Action of Disulfiram as a Modulator of the Multidrug Resistance-Linked ATP Binding Cassette Transporters MDR1 (ABCB1) and MRP1 (ABCC1)

Reactivity of zinc finger cysteines: Chemical modifications within labile zinc fingers in estrogen receptor

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