How can native mass spectrometry contribute to characterization of biomacromolecular higher-order structure and interactions?

Tong, Wenjun; Wang, Guanbo

doi:10.1016/j.ymeth.2018.04.025

Cited by 35 publications

(40 citation statements)

References 220 publications

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“…To better understand the initial stages of self‐association and aggregation of NCBD, we used electrospray ionization mass spectrometry (ESI‐MS) under native‐like conditions. This technique enables the association/dissociation of intact proteins to be monitored and provides structural insights into the corresponding protein complexes by determining their stoichiometry, topology and dynamics . In our case, the l ‐ and d ‐NCBD enantiomers as well as the racemic mixture were separately analyzed at different concentrations (10, 150 and 300 μ m ).…”

Section: Resultsmentioning

confidence: 99%

Aggregation and Amyloidogenicity of the Nuclear Coactivator Binding Domain of CREB‐Binding Protein

García

Giorgiutti

Khoury

et al. 2020

Chemistry A European J

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The nuclear coactivator binding domain (NCBD) of transcriptional co-regulator CREB-binding protein (CBP) is an example of conformationallym alleable proteins that can bind to structurally unrelated protein targets and adopt distinct folds in the respective protein complexes. Here, we show that the folding landscape of NCBD contains an alternative pathwayt hat resultsi np rotein aggregation and selfassembly into amyloid fibers. The initial steps of such protein misfolding are driven by intermolecular interactions of its N-terminal a-helix bringing multiple NCBD molecules into contact. These oligomers then undergo slow but progressive interconversion into b-sheet-containing aggregates.T o reveal the concealed aggregation potentialo fN CBD we used ac hemically synthesized mirror-image d-NCBD form. The addition of d-NCBD promoted self-assembly into amyloid precipitates presumably due to formation of thermodynamically more stable racemic b-sheet structures. The unexpecteda ggregation of NCBD needs to be taken into considerationg iven the multitude of protein-protein interactions and resulting biological functions mediated by CBP.

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Section: Resultsmentioning

confidence: 99%

Aggregation and Amyloidogenicity of the Nuclear Coactivator Binding Domain of CREB‐Binding Protein

García

Giorgiutti

Khoury

et al. 2020

Chemistry A European J

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show abstract

“…3 This work can be greatly facilitated by analytical methods capable of providing detailed information on the drug candidates' interactions with their therapeutic targets, and their ability to disrupt the molecular processes that are critical for the SARS-CoV-2 lifecycle. Native mass spectrometry (MS) has been steadily gaining popularity in the field of drug discovery, 22,23 but its applications are frequently limited to relatively homogeneous systems. Unfortunately, the large size and the extensive glycosylation of the proteins involved in the SARS-CoV-2 docking to the host cell surface (fourteen N-glycans within the ectodomain of ACE2 and at least eighteen O-and N-glycans within the S-protein ectodomain, 24 including three in its receptor binding domain, RBD) make the straightforward application of native MS to study ACE2/S-protein association challenging.…”

Section: Introductionmentioning

confidence: 99%

The utility of native MS for understanding the mechanism of action of repurposed therapeutics in COVID-19: heparin as a disruptor of the SARS-CoV-2 interaction with its host cell receptor

Yang

Kaltashov

2020

Preprint

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The emergence and rapid proliferation of the novel coronavirus (SARS-CoV-2) resulted in a global pandemic, with over six million cases and nearly four hundred thousand deaths reported worldwide by the end of May 2020. A rush to find the cures prompted re-evaluation of a range of existing therapeutics vis-à-vis their potential role in treating COVID-19, placing a premium on analytical tools capable of supporting such efforts. Native mass spectrometry (MS) has long been a tool of choice in supporting the mechanistic studies of drug/therapeutic target interactions, but its applications remain limited in the cases that involve systems with a high level of structural heterogeneity. Both SARS-CoV-2 spike protein (S-protein), a critical element of the viral entry to the host cell, and ACE2, its docking site on the host cell surface, are extensively glycosylated, making them challenging targets for native MS. However, supplementing native MS with a gas-phase ion manipulation technique (limited charge reduction) allows meaningful information to be obtained on the non-covalent complexes formed by ACE2 and the receptor-binding domain (RBD) of the S-protein. Using this technique in combination with molecular modeling also allows the role of heparin in destabilizing the ACE2/RBD association to be studied, providing critical information for understanding the molecular mechanism of its interference with the virus docking to the host cell receptor. Both short (pentasaccharide) and relatively long (eicosasaccharide) heparin oligomers form 1:1 complexes with RBD, indicating the presence of a single binding site. This association alters the protein conformation (to maximize the contiguous patch of the positive charge on the RBD surface), resulting in a notable decrease of its ability to associate with ACE2. The destabilizing effect of heparin is more pronounced in the case of the longer chains due to the electrostatic repulsion between the low-pI ACE2 and the heparin segments not accommodated on the RBD surface. In addition to providing important mechanistic information on attenuation of the ACE2/RBD association by heparin, the study demonstrates the yet untapped potential of native MS coupled to gas-phase ion chemistry as a means of facilitating rational repurposing of the existing medicines for treating COVID-19.

show abstract

“… 3 This work can be greatly facilitated by analytical methods capable of providing detailed information on the drug candidates’ interactions with their therapeutic targets and their ability to disrupt the molecular processes that are critical for the SARS-CoV-2 lifecycle. Native mass spectrometry (MS) has been steadily gaining popularity in the field of drug discovery, 22 , 23 but its applications are frequently limited to relatively homogeneous systems. Unfortunately, the large size and the extensive glycosylation of the proteins involved in the SARS-CoV-2 docking to the host cell surface (14 N-glycans within the ectodomain of ACE2 and at least 18 O- and N-glycans within the S-protein ectodomain, 24 including three in its receptor binding domain, RBD) make the straightforward application of native MS to study ACE2/S-protein association challenging.…”

mentioning

confidence: 99%

The Utility of Native MS for Understanding the Mechanism of Action of Repurposed Therapeutics in COVID-19: Heparin as a Disruptor of the SARS-CoV-2 Interaction with Its Host Cell Receptor

Yang

Kaltashov

2020

Anal. Chem.

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The emergence and rapid proliferation of the novel coronavirus (SARS-CoV-2) resulted in a global pandemic, with over 6,000,000 cases and nearly 400,000 deaths reported worldwide by the end of May 2020. A rush to find a cure prompted re-evaluation of a range of existing therapeutics vis-à-vis their potential role in treating COVID-19, placing a premium on analytical tools capable of supporting such efforts. Native mass spectrometry (MS) has long been a tool of choice in supporting the mechanistic studies of drug/therapeutic target interactions, but its applications remain limited in the cases that involve systems with a high level of structural heterogeneity. Both SARS-CoV-2 spike protein (S-protein), a critical element of the viral entry to the host cell, and ACE2, its docking site on the host cell surface, are extensively glycosylated, making them challenging targets for native MS. However, supplementing native MS with a gas-phase ion manipulation technique (limited charge reduction) allows meaningful information to be obtained on the noncovalent complexes formed by ACE2 and the receptor-binding domain (RBD) of the S-protein. Using this technique in combination with molecular modeling also allows the role of heparin in destabilizing the ACE2/RBD association to be studied, providing critical information for understanding the molecular mechanism of its interference with the virus docking to the host cell receptor. Both short (pentasaccharide) and relatively long (eicosasaccharide) heparin oligomers form 1:1 complexes with RBD, indicating the presence of a single binding site. This association alters the protein conformation (to maximize the contiguous patch of the positive charge on the RBD surface), resulting in a notable decrease in its ability to associate with ACE2. The destabilizing effect of heparin is more pronounced in the case of the longer chains due to the electrostatic repulsion between the low-p I ACE2 and the heparin segments not accommodated on the RBD surface. In addition to providing important mechanistic information on attenuation of the ACE2/RBD association by heparin, the study demonstrates the yet untapped potential of native MS coupled to gas-phase ion chemistry as a means of facilitating rational repurposing of the existing medicines for treating COVID-19.

show abstract

How can native mass spectrometry contribute to characterization of biomacromolecular higher-order structure and interactions?

Cited by 35 publications

References 220 publications

Aggregation and Amyloidogenicity of the Nuclear Coactivator Binding Domain of CREB‐Binding Protein

Aggregation and Amyloidogenicity of the Nuclear Coactivator Binding Domain of CREB‐Binding Protein

The utility of native MS for understanding the mechanism of action of repurposed therapeutics in COVID-19: heparin as a disruptor of the SARS-CoV-2 interaction with its host cell receptor

The Utility of Native MS for Understanding the Mechanism of Action of Repurposed Therapeutics in COVID-19: Heparin as a Disruptor of the SARS-CoV-2 Interaction with Its Host Cell Receptor

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