Temperature‐Controlled Electrospray Ionization: Recent Progress and Applications

Harrison, Julian A.; Pruška, Adam; Oganesyan, Irina; Bittner, Philipp; Zenobi, Renato

doi:10.1002/chem.202102474

Cited by 14 publications

(6 citation statements)

References 99 publications

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“…This temperature-induced effect for System 1 is consistent with a known phenomenon in mass spectrometry, where excessive source temperatures promote fragmentation and denaturation. 43,44 When we applied this temperature ramp to System 2, it became evident that detection of the n = 4 ion was virtually non-existent, even at low source temperatures. This very low abundance of highcoordinate ions suggests that other aspects of the instrument design, rather than in-source thermal ion fragmentation, are the cause of softness limitations in System 2.…”

Section: Impact Of Source Temperatures On Ion Fragmentation Profilesmentioning

confidence: 99%

How soft is your ESI-MS anyway?

Chagunda,

Williams,

Fisher

et al. 2024

Preprint

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Electrospray ionization mass spectrometry (ESI-MS) is a powerful tool for identifying and characterizing organometallic and coordination compounds. However, detection of fragile structures bound by weaker intermolecular forces can be significantly limited in ESI-MS owing to the use of relatively harsh instrument conditions and configurations. In this study, a set of tests was developed to assess the softness of ESI-MS systems. Two variants are presented: positive ion mode, utilizing a mixture of sodium ions and triphenylphosphine oxide producing [Na(OPPh3)n]+ ions (n = 1-4), and negative ion mode utilizing Pd(PPh3)4 and sulfonated triphenylphosphine producing [Pd(L)(PPh3)n]– ions (n = 0-2), where softer instrument conditions preserve a higher proportion of the high-coordinate ions and harsher conditions will result in increased detection of products of ion fragmentation. The results revealed notable variations in instrument softness, which were influenced by a combination of instrument design and experimental parameters. Meticulously optimizing experimental conditions and ESI-MS parameters is essential to achieving the softest ionization possible, ensuring reliable analysis where applicable. This study offers valuable insight through straightforward tests that can be employed to assess the suitability of an instrument for specific research needs.

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Section: Impact Of Source Temperatures On Ion Fragmentation Profilesmentioning

confidence: 99%

How soft is your ESI-MS anyway?

Chagunda,

Williams,

Fisher

et al. 2024

Preprint

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“…In agreement with principles of enzyme mechanisms, previous MS studies have highlighted the importance of controlling temperature. 37,38 This is of particular importance for measuring PLA 2 activity, as temperature can affect both enzyme stability and lipid membrane conformation/integrity. The effect of temperature on PDx enzymatic activity was investigated under the standard assay conditions (PDx (6 nM); DOPC liposomes (80 μM), in 100 mM NH 4 OAc, 2 mM in Ca(OAc) 2 ).…”

Section: Effect Of Lpc Concentration On the Ionization Of Dopcmentioning

confidence: 99%

Continuous Electrospray Ionization Mass Spectrometry Assay for Measuring Phospholipase Activity against Liposomes

Harrison

Kelso

Beck

2023

J. Am. Soc. Mass Spectrom.

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Phospholipases have diverse roles in lipid and cell membrane biology. In animal venoms, they can have roles as neurotoxins or myotoxins that disrupt the integrity of cell membranes. In this work, we describe a temperature-controlled, continuous electrospray ionization mass spectrometry (ESI-MS) assay for measuring phospholipase A 2 activity against liposomes. The enzyme used in this assay was paradoxin, which is a neurotoxic trimeric phospholipase A 2 from inland taipan snake venom. Previously developed ESI-MS-based phospholipase assays have been discontinuous and analyzed hydrolysis of single lipid molecules by liquid chromatography ESI-MS. In this work, a continuous assay was developed against liposomes, a more complex substrate that more closely reflects the natural substrate for paradoxin. The assay confirmed the requirement for Ca 2+ and allowed measurement of Michaelis−Menten-type parameters. The use of ESI-MS for lipid detection enabled nuanced insights into the effect of changing assay conditions not only on the enzyme but also on the liposome substrate. Changing the metal ion concentrations did not significantly change the liposomes but did affect enzymatic activity. Increasing temperature did not substantially affect the secondary structure of paradoxin but affected liposome size, resulting in increased enzymatic activity consistent with the disruption of the phosphatidylcholine membrane, increasing accessibility of sn-2 ester bonds. The continuous ESI-MS method described herein can be applied to other enzyme reactions, particularly those which utilize complex lipid substrates.

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“…Multiple designs exist for TC-nESI-MS sources, each of which has been used to investigate a range of biomolecules. 30 However, this technique has yet to be employed for proteomics analysis.…”

Section: ■ Introductionmentioning

confidence: 99%

“…The resulting temperature-induced changes in a biomolecule can then be detected by MS. Multiple designs exist for TC-nESI-MS sources, each of which has been used to investigate a range of biomolecules . However, this technique has yet to be employed for proteomics analysis.…”

Section: Introductionmentioning

confidence: 99%

Streamlining the Analysis of Proteins from Snake Venom

Oganesyan,

Jenkins,

Laustsen

et al. 2024

J. Proteome Res.

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Investigating snake venom is necessary for developing new treatments for envenoming and harnessing the therapeutic potential that lies within venom toxins. Despite considerable efforts in previous research, several technical challenges remain for characterizing the individual components within such complex mixtures. Here, we present native and top-down mass spectrometry (MS) workflows that enable the analysis of individual venom proteins within complex mixtures and showcase the utility of these methodologies on King cobra (Ophiophagus hannah) venom. First, we coupled ion mobility spectrometry for separation and electron capture dissociation for charge reduction to resolve highly convoluted mass spectra containing multiple proteins with masses ranging from 55 to 127 kDa. Next, we performed a top-down glycomic analysis of a 25.5 kDa toxin, showing that this protein contains a fucosylated complex glycan. Finally, temperature-controlled nanoelectrospray mass spectrometry facilitated the top-down sequence analysis of a β-cardiotoxin, which cannot be fragmented by collisional energy due to its disulfide bond pattern. The work presented here demonstrates the applicability of new and promising MS methods for snake venom analysis.

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Temperature‐Controlled Electrospray Ionization: Recent Progress and Applications

Cited by 14 publications

References 99 publications

How soft is your ESI-MS anyway?

How soft is your ESI-MS anyway?

Continuous Electrospray Ionization Mass Spectrometry Assay for Measuring Phospholipase Activity against Liposomes

Streamlining the Analysis of Proteins from Snake Venom

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