Fingerprinting the tertiary structure of electroadsorbed lysozyme at soft interfaces by electrostatic spray ionization mass spectrometry

Eulate, Eva Alvárez de; Qiao, Liang; Scanlon, Micheál D.; Arrigan, Damien W. M.

doi:10.1039/c4cc05545d

Cited by 26 publications

(42 citation statements)

References 26 publications

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“…However, protein−protein interactions, which are likely to play a key role in behaviour at the oil−water interface, and therefore the impact of interfacial concentration of adsorbed protein have not been addressed. Recently, the method of adsorptive stripping voltammetry (AdSV) was applied to achieve low limits of detection for lysozyme at an array of micrometre‐sized ITIES (μITIES array) and subsequently combined with electrostatic spray ionisation‐mass spectrometry (ESTASI‐MS) to analyse protein structural changes . The AdSV method effectively pre‐concentrates the protein at the interface via a potential‐controlled electroadsorption for a fixed time, providing an enhanced current response to the protein during the subsequent desorption step.…”

Section: Introductionsupporting

confidence: 67%

“…Cyt c is known to form dimers, trimers, tetramers and higher order oligomers under certain conditions; perhaps the combination of acidic aqueous phase, interfacial complexation of organic electrolyte anion, and adsorption of the complex at the interface leads to conditions that promote such oligomerisation. Previous work illustrated the partial unfolding of lysozyme at soft interfaces (w/gelled‐DCH) when this protein was electrochemically adsorbed . Cyt c can undergo more extensive unfolding as it is less stabilised by disulfide bonds than lysozyme.…”

Section: Resultsmentioning

confidence: 99%

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Electrochemically Induced Formation of Cytochrome c Oligomers at Soft Interfaces

2017

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The formation of cytochrome c oligomers was induced at liquid−gel and liquid−liquid interfaces via electroadsorption. At an optimum interfacial potential (Eads=0.975 V), the protein was accumulated at these soft interfaces. It was found that as the concentration of adsorbed protein increased, a single voltammetric peak evolved into double and triple peaks (tads=300 s). Analysis of the protein that accumulated at the interfaces by polyacrylamide gel electrophoresis indicated the presence of oligomeric species, corresponding to dimers (ca. 27 kD), trimers (ca. 35 kD), and even larger species (>250 kD) after prolonged electroadsorption (tads=2 h) at macro‐scale soft interfaces. Accordingly, it was possible to electrochemically induce oligomerisation at these soft interfaces, which can be tuned through experimental factors such as interfacial potential difference, electroadsorption time, and bulk solution concentration. These results suggest the use of electrochemistry at soft interfaces as a strategy for the investigation of protein oligomerisation and its inhibition.

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Section: Introductionsupporting

confidence: 67%

Section: Resultsmentioning

confidence: 99%

Electrochemically Induced Formation of Cytochrome c Oligomers at Soft Interfaces

2017

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“…The catalytic aspartic acid residues (D70) of the C‐type lysozyme ( Gallus gallus ) (Alvarez de Eulate et al . ) and the catalytic threonine residues (T50) of the lysozyme of E. coli (Mercer et al . ) are conserved in the YeaZ protein of V. harveyi (D 112 and T 71).…”

Section: Discussionmentioning

confidence: 99%

Promoting resuscitation of viable but nonculturable cells ofVibrio harveyiby a resuscitation-promoting factor-like protein YeaZ

Chen

zhao

et al. 2016

J Appl Microbiol

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“…76) e principle of this contactless spray was proposed from a capacitive coupling e ect. is technique termed as electrostatic-spray ionization has been used for analysis of adsorbed proteins, 77) perfumes, 78) and imaging applications.…”

Section: Polarizationmentioning

confidence: 99%

Electrospray Modifications for Advancing Mass Spectrometric Analysis

Meher

Chen

2017

Mass Spectrometry

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Generation of analyte ions in gas phase is a primary requirement for mass spectrometric analysis. One of the ionization techniques that can be used to generate gas phase ions is electrospray ionization (ESI). ESI is a so ionization method that can be used to analyze analytes ranging from small organics to large biomolecules. Numerous ionization techniques derived from ESI have been reported in the past two decades.ese ion sources are aimed to achieve simplicity and ease of operation. Many of these ionization methods allow the exibility for elimination or minimization of sample preparation steps prior to mass spectrometric analysis. Such ion sources have opened up new possibilities for taking scienti c challenges, which might be limited by the conventional ESI technique. us, the number of ESI variants continues to increase. is review provides an overview of ionization techniques based on the use of electrospray reported in recent years. Also, a brief discussion on the instrumentation, underlying processes, and selected applications is also presented.

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Fingerprinting the tertiary structure of electroadsorbed lysozyme at soft interfaces by electrostatic spray ionization mass spectrometry

Cited by 26 publications

References 26 publications

Electrochemically Induced Formation of Cytochrome c Oligomers at Soft Interfaces

Electrochemically Induced Formation of Cytochrome c Oligomers at Soft Interfaces

Promoting resuscitation of viable but nonculturable cells ofVibrio harveyiby a resuscitation-promoting factor-like protein YeaZ

Electrospray Modifications for Advancing Mass Spectrometric Analysis

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