Investigation on the labile interactions of proteins and ligands using electrospray ionization mass spectrometry combined with a mathematical method

Tang, Jun; Fu, Qiang; Cui, Meng; Liu, Zhiqiang; Liu, Shuying

doi:10.1002/jms.3785

Cited by 4 publications

(7 citation statements)

References 20 publications

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“…Studies have shown that some complexes stabilized by hydrophobic interactions or ionic interactions in solutions were not stable during ionization. The in-source partial or complete dissociation of these PL complexes happened even under “gentle” ESI ionization conditions. − …”

Section: Introductionmentioning

confidence: 99%

“…Many strategies have been developed to avoid the in-source dissociation, such as hydrogen/deuterium exchange, affinity chromatography, cold spray ionization, competitive experiments, and diffusion measurements. − The researchers also found that the introduction of some additives into PL solution could increase the survival probability of the labile PL complexes. ,− However, it is not easy to find a suitable additive to protect the labile PL complexes. Until now, only several effective additives have been found.…”

Section: Introductionmentioning

confidence: 99%

“…Lysozyme is a globular protein and is widely distributed in various biological fluids and tissues. It has many pharmaceutical functions, such as antiviral, anti-inflammatory, antitumor activities, and so on. , It is known to combine with many active small molecules for therapeutic applications. , Many flavonoids binding to lysozyme have been reported, such as rutin, luteolin, etc. ,, …”

Section: Introductionmentioning

confidence: 99%

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Stabilization of Labile Lysozyme–Ligand Interactions in Native Electrospray Ionization Mass Spectrometry

Zhao

Xing

et al. 2023

J. Am. Soc. Mass Spectrom.

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Flavonoids are polyphenolic secondary metabolites with extensive biological activities and pharmacological effects. Exploring the interactions of flavonoids with proteins may be helpful for understanding their biological processes. Electrospray ionization mass spectrometry (ESI-MS) is a powerful tool to characterize the noncovalent protein−ligand (PL) complexes. However, some protein−flavonoid complexes are labile during electrospray ionization. Here, the labile lysozyme−flavonoid (rutin, icariin, and naringin) complexes were determined by direct ESI-MS without derivation. It has been found that low amounts of N-methylpyrrolidinone and dimethylformamide can protect labile lysozyme-flavonoid complexes away from dissociation during electrospray ionization process. The intact lysozyme-flavonoid complexes were specifically observed in mass spectra, and the measured binding affinities by ESI-MS were matched with the fluorescence data. The effects of additives on the analysis of lysozymeflavonoid complexes were investigated by ESI-MS, combined with the molecular docking and fluorescence. This strategy was helpful to investigate the labile PL interactions by direct ESI-MS.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Stabilization of Labile Lysozyme–Ligand Interactions in Native Electrospray Ionization Mass Spectrometry

Zhao

Xing

et al. 2023

J. Am. Soc. Mass Spectrom.

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“…A number of analytical techniques are available for the characterization of binding behavior of proteins with ligands, most of which are complex, costly and time‐consuming. Many of them need complicated analyte preparation such as X‐ray analysis, nuclear magnetic resonance‐, mass‐ and circular dichroism (CD) spectrometry . Among all analytical techniques, affinity capillary electrophoresis (ACE) has prevailed as a fast, precise and parsimonious method for binding studies between proteins and various ligands in the last years .…”

Section: Introductionmentioning

confidence: 99%

Interaction of albumins and heparinoids investigated by affinity capillary electrophoresis and free flow electrophoresis

et al. 2017

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A fast and precise affinity capillary electrophoresis (ACE) method has been applied to investigate the interactions between two serum albumins (HSA and BSA) and heparinoids. Furthermore, different free flow electrophoresis methods were developed to separate the species which appears owing to interaction of albumins with pentosan polysulfate sodium (PPS) under different experimental conditions. For ACE experiments, the normalized mobility ratios (∆R/R ), which provided information about the binding strength and the overall charge of the protein-ligand complex, were used to evaluate the binding affinities. ACE experiments were performed at two different temperatures (23 and 37°C). Both BSA and HSA interact more strongly with PPS than with unfractionated and low molecular weight heparins. For PPS, the interactions can already be observed at low mg/L concentrations (3 mg/L), and saturation is already obtained at approximately 20 mg/L. Unfractionated heparin showed almost no interactions with BSA at 23°C, but weak interactions at 37°C at higher heparin concentrations. The additional signals also appeared at higher concentrations at 37°C. Nevertheless, in most cases the binding data were similar at both temperatures. Furthermore, HSA showed a characteristic splitting in two peaks especially after interacting with PPS, which is probably attributable to the formation of two species or conformational change of HSA after interacting with PPS. The free flow electrophoresis methods have confirmed and completed the ACE experiments.

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“…The investigation of binding behavior between proteins and ligands is an essential field of biochemistry and also the basis of drug development . Various analytical techniques are well suited to study protein–ligand interactions, especially X‐ray analysis of cocrystallized proteins, nuclear magnetic resonance‐, mass‐ and circular dichroism (CD) spectroscopy . Almost all of these techniques require pure proteins and partly large amounts of samples.…”

Section: Introductionmentioning

confidence: 99%

Using affinity capillary electrophoresis and computational models for binding studies of heparinoids with p‐selectin and other proteins

et al. 2017

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A fast and precise affinity capillary electrophoresis (ACE) method has been developed and applied for the investigation of the binding interactions between P-selectin and heparinoids as potential P-selectin inhibitors in the presence and absence of calcium ions. Furthermore, model proteins and vitronectin were used to appraise the binding behavior of P-selectin. The normalized mobility ratios (∆R/R ), which provided information about the binding strength and the overall charge of the protein-ligand complex, were used to evaluate the binding affinities. It was found that P-selectin interacts more strongly with heparinoids in the presence of calcium ions. P-selectin was affected by heparinoids at the concentration of 3 mg/L. In addition, the results of the ACE experiments showed that among other investigated proteins, albumins and vitronectin exhibited strong interactions with heparinoids. Especially with P-selectin and vitronectin, the interaction may additionally induce conformational changes. Subsequently, computational models were applied to interpret the ACE experiments. Docking experiments explained that the binding of heparinoids on P-selectin is promoted by calcium ions. These docking models proved to be particularly well suited to investigate the interaction of charged compounds, and are therefore complementary to ACE experiments.

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Investigation on the labile interactions of proteins and ligands using electrospray ionization mass spectrometry combined with a mathematical method

Cited by 4 publications

References 20 publications

Stabilization of Labile Lysozyme–Ligand Interactions in Native Electrospray Ionization Mass Spectrometry

Stabilization of Labile Lysozyme–Ligand Interactions in Native Electrospray Ionization Mass Spectrometry

Interaction of albumins and heparinoids investigated by affinity capillary electrophoresis and free flow electrophoresis

Using affinity capillary electrophoresis and computational models for binding studies of heparinoids with p‐selectin and other proteins

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