Native ion mobility mass spectrometry reveals that small organic acid fragments impart gas‐phase stability to carbonic anhydrase II

Veale, Clinton G. L.; Jimenez, Maria Mateos; Mackay, C. Logan; Clarke, David J.

doi:10.1002/rcm.8570

Cited by 14 publications

(14 citation statements)

References 32 publications

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“…Following the commercialization of quadrupole/TWIMS/time-of-flight mass spectrometers 4 , modern technological developments have further extended the range of CIU examples. Current applications of CIU encompass probing protein-protein complexes 12,13 , protein-ligand interactions 9,[14][15][16][17][18] , and binding of lipids to membrane proteins 19,20 . CIU also plays a key role to investigate therapeutically-relevant proteins, allowing more conformational insights into monoclonal antibodies (mAbs) [21][22][23][24] , biosimilars 25 and antibody-drugconjugates 26,27 .…”

Section: Introductionmentioning

confidence: 99%

“…Following the commercialization of quadrupole/TWIMS/time-of-flight mass spectrometers, modern technological developments have further extended the range of CIU examples. Current applications of CIU encompass probing protein–protein complexes, , protein–ligand interactions, ,− and binding of lipids to membrane proteins. , CIU also plays a key role to investigate therapeutically relevant proteins, allowing more conformational insights into monoclonal antibodies (mAbs), − biosimilars, and antibody–drug conjugates. , In particular, Tian et al demonstrated that subtle differences between intact mAbs subclasses, with different numbers and patterns of disulfide bridges, could be distinguished based on their gas-phase unfolding. Recently, Botzanowski et al expanded this strategy to the middle-level classification of mAbs subclasses, which provides a more clear-cut categorization than the intact level …”

Section: Introductionmentioning

confidence: 99%

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Toward Automation of Collision-Induced Unfolding Experiments through Online Size Exclusion Chromatography Coupled to Native Mass Spectrometry

et al. 2020

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Ion mobility-based collision induced unfolding (CIU) has gained increasing attention to probe gas-phase unfolding of proteins and their noncovalent complexes, notably for biotherapeutics. CIU detects subtle conformational changes of proteins and emerges as an attractive alternative to circumvent poor IM resolution. However, CIU still lacks in automation for buffer exchange and data acquisition, precluding its wide adoption. We present here an automated workflow for CIU experiments, from sample preparation to data interpretation using online size exclusion chromatography coupled to native ion mobility mass spectometry (SEC-CIU). Online automated SEC-CIU experiments offer several benefits over nanoESI-CIU, among which i) improved and fast desalting compared to manual buffer exchange used for classical CIU experiments; ii) drastic reduction of the overall data collection time process along with iii) maintaining the number of unfolding transitions. We then evaluate the potential of SEC-CIU to distinguish monoclonal antibodies (mAbs) subclasses, illustrating the efficiency of our method for rapid mAb subclass identification at both intact and middle levels. Finally, we demonstrate that CIU data acquisition time can be further reduced either by setting up a scheduled CIU method relying on diagnostic trap collision voltages or by implementing mAbs-multiplexed SEC-CIU analyses to maximize information content in a single experiment. Altogether, our results confirm the suitability of SEC-CIU to automate CIU experiments, particularly for the fast characterization of next generation mAb-based products.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Toward Automation of Collision-Induced Unfolding Experiments through Online Size Exclusion Chromatography Coupled to Native Mass Spectrometry

et al. 2020

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“…1,2 Until now, native electrospray ionization mass spectrometry (native ESI-MS) has emerged as a powerful analytical approach to probe the protein-ligand (P-L) interactions. 3,4 This method employs the soft ionization technique ESI to transfer the intact P-L complexes from the liquid phase to the gas phase to conserve the weak non-covalent interaction. 5 P-L complexes with millimolar dissociation constants (K d ) can be detected.…”

Section: Introductionmentioning

confidence: 99%

Native electrospray ionization mass spectrometry combined with molecular docking for the characterization of ginsenoside–myoglobin interactions

Zhao

Zheng

et al. 2021

Rapid Comm Mass Spectrometry

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Rationale: The interactions between proteins and ligands are involved in many biological processes and early stages of drug development. Native electrospray ionization mass spectrometry (native ESI-MS) has played an important role in the characterization of protein-ligand interactions. Herein, native ESI-MS combined with molecular docking was used for the characterization of ginsenoside-myoglobin (Mb) interactions. Methods: The binding of ginsenosides (Rb 3 , Rc, Rd, Re) to Mb was determined by native ESI-MS. Titration experiments were performed for the calculation of the dissociation constants (K d ) of the complexes. Molecular docking was used to simulate the binding of ginsenosides with Mb by AutoDock.Results: The ginsenoside-Mb complex with stoichiometric ratio 1:1 was observed by native ESI-MS. The K d values determined by the direct calculation method were matched with those obtained by the curve fitting method. However, the relative standard deviations (RSDs) obtained by direct calculation were larger than those obtained by curve fitting. From the molecular docking, it was inferred that hydrophobic interactions, hydrogen bonding and Van der Waals forces participate in the binding of ginsenosides to proteins. Conclusions:The ginsenoside-Mb interactions can be characterized by ESI-MS combined with molecular docking. This approach can be helpful to investigate the interactions between natural drugs and proteins in various diseases.

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“…[7] Whilst conventionally underutilised, the speed and sensitivity inherent to native mass spectrometry (MS) has seen it emerge as a powerful fragment screening technique, which is capable of detecting weak interactions between unlabelled and untethered small molecular fragments and target proteins in the gas phase. [8][9][10][11] MS ligand binding experiments typically look to detect a molecular ion correlating to the apo protein in its native state (M) with the emergence of a new ion corresponding to the deconvoluted molecular mass of the proteinligand complex (M•L) indicating ligand binding. Comparison of the relative intensities of these ion peaks provides a rough indication of binding strength.…”

Section: Introductionmentioning

confidence: 99%

Native mass spectrometry guided screening identifies hit fragments for HOP-HSP90 PPI inhibition

Veale

Vaaltyn

Mateos-Jimenez

et al. 2021

Preprint

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Contemporary medicinal chemistry considers fragment-based drug discovery (FBDD) and inhibition of Protein-Protein Interactions (PPI), as important means of expanding druggable chemical space. However, the ability to robustly identify valid fragments and PPI inhibitors is an enormous challenge, requiring the application of sensitive biophysical methodology. Accordingly, in this study, we exploited the speed and sensitivity of nano-ESI native mass spectrometry to identify a small collection of fragments, which bind to the TPR2AB domain of HOP. Further biological assessment of a small selection of binding fragments showed that this binding translated into PPI inhibitory activity between the TPR2A domain of HOP and the HSP90- C terminal domain. An in silico assessment of binding fragments, at the PPI interfacial region provided valuable structural insight for future fragment elaboration strategies, including the identification of losartan as a weak, albeit dose dependent inhibitor of the target PPI.

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Native ion mobility mass spectrometry reveals that small organic acid fragments impart gas‐phase stability to carbonic anhydrase II

Abstract: Ion Mobility Mass Spectrometry (IMMS) reveals that small organic acid fragments impart gasphase stability to carbonic anhydrase II', Rapid communications in mass spectrometry.

Cited by 14 publications

References 32 publications

Toward Automation of Collision-Induced Unfolding Experiments through Online Size Exclusion Chromatography Coupled to Native Mass Spectrometry

Toward Automation of Collision-Induced Unfolding Experiments through Online Size Exclusion Chromatography Coupled to Native Mass Spectrometry

Native electrospray ionization mass spectrometry combined with molecular docking for the characterization of ginsenoside–myoglobin interactions

Native mass spectrometry guided screening identifies hit fragments for HOP-HSP90 PPI inhibition

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