Native mass spectrometry of human carbonic anhydrase I and its inhibitor complexes

Zoppi, Carlotta; Nocentini, Alessio; Pratesi, Alessandro; Messori, Luigi

doi:10.1007/s00775-020-01818-8

Cited by 5 publications

(10 citation statements)

References 90 publications

(117 reference statements)

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“…For these experiments, the proteins were treated in physiological-like conditions, using a well-established protocol [ 18 , 36 , 37 ] that can assure a protein reactivity as similar as possible to that of the physiological environment. Moreover, the spectra were acquired with a methodology able to preserve the protein in its native state, conserving unaltered all the covalent interactions formed in solution and most of the non-covalent ones [ 38 ]. From the inspection of the obtained spectra, clearly emerged two different reactivity patterns depending on the nature of the biomolecules.…”

Section: Resultsmentioning

confidence: 99%

Thermodynamic Evaluation of the Interactions between Anticancer Pt(II) Complexes and Model Proteins

et al. 2021

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In this work, we have analysed the binding of the Pt(II) complexes ([PtCl(4′-phenyl-2,2′:6′,2″-terpyridine)](CF3SO3) (1), [PtI(4′-phenyl-2,2′:6′,2″-terpyridine)](CF3SO3) (2) and [PtCl(1,3-di(2-pyridyl)benzene) (3)] with selected model proteins (hen egg-white lysozyme, HEWL, and ribonuclease A, RNase A). Platinum coordination compounds are intensively studied to develop improved anticancer agents. In this regard, a critical issue is the possible role of Pt-protein interactions in their mechanisms of action. Multiple techniques such as differential scanning calorimetry (DSC), electrospray ionization mass spectrometry (ESI-MS) and UV-Vis absorbance titrations were used to enlighten the details of the binding to the different biosubstrates. On the one hand, it may be concluded that the affinity of 3 for the proteins is low. On the other hand, 1 and 2 strongly bind them, but with major binding mode differences when switching from HEWL to RNase A. Both 1 and 2 bind to HEWL with a non-specific (DSC) and non-covalent (ESI-MS) binding mode, dominated by a 1:1 binding stoichiometry (UV-Vis). ESI-MS data indicate a protein-driven chloride loss that does not convert into a covalent bond, likely due to the unfavourable complexes’ geometries and steric hindrance. This result, together with the significant changes of the absorbance profiles of the complex upon interaction, suggest an electrostatic binding mode supported by some stacking interaction of the aromatic ligand. Very differently, in the case of RNase A, slow formation of covalent adducts occurs (DSC, ESI-MS). The reactivity is higher for the iodo-compound 2, in agreement with iodine lability higher than chlorine.

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Section: Resultsmentioning

confidence: 99%

Thermodynamic Evaluation of the Interactions between Anticancer Pt(II) Complexes and Model Proteins

et al. 2021

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“…Respective ESI mass spectra were acquired through direct infusion at 7 μL min −1 flow rate. The samples were prepared in LC-MS grade solvents, following a well-established protocol previously developed [ 29 , 30 , 35 , 69 ]. To promote the ionization in positive mode, 0.1% v / v of formic acid was added just before infusion.…”

Section: Methodsmentioning

confidence: 99%

On the Different Mode of Action of Au(I)/Ag(I)-NHC Bis-Anthracenyl Complexes Towards Selected Target Biomolecules

et al. 2020

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Gold and silver N-heterocyclic carbenes (NHCs) are emerging for therapeutic applications. Multiple techniques are here used to unveil the mechanistic details of the binding to different biosubstrates of bis(1-(anthracen-9-ylmethyl)-3-ethylimidazol-2-ylidene) silver chloride [Ag(EIA)2]Cl and bis(1-(anthracen-9-ylmethyl)-3-ethylimidazol-2-ylidene) gold chloride [Au(EIA)2]Cl. As the biosubstrates, we tested natural double-stranded DNA, synthetic RNA polynucleotides (single-poly(A), double-poly(A)poly(U) and triple-stranded poly(A)2poly(U)), DNA G-quadruplex structures (G4s), and bovine serum albumin (BSA) protein. Absorbance and fluorescence titrations, mass spectrometry together with melting and viscometry tests show significant differences in the binding features between silver and gold compounds. [Au(EIA)2]Cl covalently binds BSA. It is here evidenced that the selectivity is high: low affinity and external binding for all polynucleotides and G4s are found. Conversely, in the case of [Ag(EIA)2]Cl, the binding to BSA is weak and relies on electrostatic interactions. [Ag(EIA)2]Cl strongly/selectively interacts only with double strands by a mechanism where intercalation plays the major role, but groove binding is also operative. The absence of an interaction with triplexes indicates the major role played by the geometrical constraints to drive the binding mode.

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“…The interaction of sulfonamide inhibitors and human CA I has been addressed recently using native mass spectrometry, which allows for preservation of non-covalent interactions during the transition of the protein–ligand complex from solution to the gas phase. In this way, stoichiometric information about the composition of the complex was obtained, and the binding of different inhibitors was clarified [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Ligation Motifs in Zinc-Bound Sulfonamide Drugs Assayed by IR Ion Spectroscopy

et al. 2022

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The sulfonamide–zinc ion interaction, performing a key role in various biological contexts, is the focus of the present study, with the aim of elucidating ligation motifs in zinc complexes of sulfa drugs, namely sulfadiazine (SDZ) and sulfathiazole (STZ), in a perturbation-free environment. To this end, an approach is exploited based on mass spectrometry coupled with infrared multiple photon dissociation (IRMPD) spectroscopy backed by quantum chemical calculations. IR spectra of Zn(H2O+SDZ−H)+ and Zn(H2O+STZ−H)+ ions are consistent with a three-coordinate zinc complex, where ZnOH+ binds to the uncharged sulfonamide via N(heterocycle) and O(sulfonyl) donor atoms. Alternative prototropic isomers Zn(OH2)(SDZ−H)+ and Zn(OH2)(STZ−H)+ lie 63 and 26 kJ mol−1 higher in free energy, respectively, relative to the ground state Zn(OH)(SDZ)+ and Zn(OH)(STZ)+ species and do not contribute to any significant extent in the sampled population.

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Native mass spectrometry of human carbonic anhydrase I and its inhibitor complexes

Cited by 5 publications

References 90 publications

Thermodynamic Evaluation of the Interactions between Anticancer Pt(II) Complexes and Model Proteins

Thermodynamic Evaluation of the Interactions between Anticancer Pt(II) Complexes and Model Proteins

On the Different Mode of Action of Au(I)/Ag(I)-NHC Bis-Anthracenyl Complexes Towards Selected Target Biomolecules

Ligation Motifs in Zinc-Bound Sulfonamide Drugs Assayed by IR Ion Spectroscopy

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