Native Desorption Electrospray Ionization Liberates Soluble and Membrane Protein Complexes from Surfaces

Ambrose, Stephen J.; Housden, Nicholas G.; Gupta, K. P.; Fan, Jieyuan; White, Paul; Yen, Hsin‐Yung; Marcoux, Julien; Kleanthous, Colin; Hopper, Jonathan T. S.; Robinson, Carol V.

doi:10.1002/anie.201704849

Cited by 50 publications

(49 citation statements)

References 33 publications

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“…The recorded spectrum is dominated by 15+ to 17+ BSA ions, which is consistent with the finding of the native DESI work from Robinson's group. 28 However, the width of ion peaks in our native BSA study is found to be broader. Apart from the differences in resolving power afforded by a Q-ToF compared to an Orbitrap mass analyzer, we suggest that the broad peak width may also be a result of the non-desalting protein sample preparation method employed in our study.…”

Section: Solvent Filtering For Native Desimentioning

confidence: 50%

Probing Folded Proteins and Intact Protein Complexes by Desorption Electrospray Ionization Mass Spectrometry

Yan¹,

Bunch²

2020

Preprint

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Native mass spectrometry (Native MS) enables the study of intact proteins as well as non-covalent protein-protein and protein-ligand complexes in their biological state. In this work we present the application of a prototype Waters DESI source for rapid surface measurements of folded and native protein structures. Ions with narrow charge state distribution (CSD), i.e. folded structures are observed in the spectra of protein samples with the molecular weight ranging from 8.6 kDa up to 66.4 kDa. Intact protein complexes of holo-myoglobin and tetrameric hemoglobin are also successfully detected from a surface. These results reveal that DESI could be gentle enough to detect compact structures and noncovalent bond interactions. We also examine whether unfolded proteins and protein complexes can refold during transient spray solvent-sample interactions during DESI. Our results from ion mobility experiments of standards of ubiquitin, cytochrome c and protein complex myoglobin indicate that such phenomenon may occur, presenting artificial native-like spectra. Nevertheless, the observation of hemoglobin tetramer is promising as it demonstrates the capability of DESI to maintain truly native structures.

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Section: Solvent Filtering For Native Desimentioning

confidence: 50%

Probing Folded Proteins and Intact Protein Complexes by Desorption Electrospray Ionization Mass Spectrometry

Yan¹,

Bunch²

2020

Preprint

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“…Analysis by high resolution MS revealed that the charge state distributions observed depend on the detergent used in the desorption plume, suggesting that on-surface detergent exchange was occurring. Moreover, specific binding to MPs could be observed by adding ligands to the desorption spray [128]. Excitingly, this opens up the possibility for the study of membraneembedded MPs on surfaces directly by DESI-MS.…”

Section: Alternative Ionization Methodsmentioning

confidence: 99%

“…2) [123,127]. Using DESI, MPs prepared in detergents and deposited onto glass surfaces could be desorbed by electrospraying a detergent-containing solution onto the surface [128]. Analysis by high resolution MS revealed that the charge state distributions observed depend on the detergent used in the desorption plume, suggesting that on-surface detergent exchange was occurring.…”

Section: Alternative Ionization Methodsmentioning

confidence: 99%

Mass spectrometry-enabled structural biology of membrane proteins

Calabrese

Radford

2018

Methods

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a b s t r a c tThe last $25 years has seen mass spectrometry (MS) emerge as an integral method in the structural biology toolkit. In particular, MS has enabled the structural characterization of proteins and protein assemblies that have been intractable by other methods, especially those that are large, heterogeneous or transient, providing experimental evidence for their structural organization in support of, and in advance of, high resolution methods. The most recent frontier conquered in the field of MS-based structural biology has been the application of established methods for studying water soluble proteins to the more challenging targets of integral membrane proteins. The power of MS in obtaining structural information has been enabled by advances in instrumentation and the development of hyphenated mass spectrometry-based methods, such as ion mobility spectrometry-MS, chemical crosslinking-MS and other chemical labelling/footprinting-MS methods. In this review we detail the insights garnered into the structural biology of membrane proteins by applying such techniques. Application and refinement of these methods has yielded unprecedented insights in many areas, including membrane protein conformation, dynamics, lipid/ligand binding, and conformational perturbations due to ligand binding, which can be challenging to study using other methods.

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“…Clearly, DESI requires further investigation to develop our understanding for protein analysis and this is certainly a barrier to its adoption in routine or high-throughput analysis environments. One study has reported DESI for native protein analysis of purified proteins, successfully ionising large non-covalent complexes such as GroEL 14-mer [69]. Insoluble membrane proteins were also analysed by the addition of detergents to the DESI solvent system.…”

Section: Desorption Techniquesmentioning

confidence: 99%

In situ mass spectrometry analysis of intact proteins and protein complexes from biological substrates

Hale

Cooper

2020

Biochemical Society Transactions

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Advances in sample preparation, ion sources and mass spectrometer technology have enabled the detection and characterisation of intact proteins. The challenges associated include an appropriately soft ionisation event, efficient transmission and detection of the often delicate macromolecules. Ambient ion sources, in particular, offer a wealth of strategies for analysis of proteins from solution environments, and directly from biological substrates. The last two decades have seen rapid development in this area. Innovations include liquid extraction surface analysis, desorption electrospray ionisation and nanospray desorption electrospray ionisation. Similarly, developments in native mass spectrometry allow protein–protein and protein–ligand complexes to be ionised and analysed. Identification and characterisation of these large ions involves a suite of hyphenated mass spectrometry techniques, often including the coupling of ion mobility spectrometry and fragmentation techniques. The latter include collision, electron and photon-induced methods, each with their own characteristics and benefits for intact protein identification. In this review, recent developments for in situ protein analysis are explored, with a focus on ion sources and tandem mass spectrometry techniques used for identification.

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Native Desorption Electrospray Ionization Liberates Soluble and Membrane Protein Complexes from Surfaces

Cited by 50 publications

References 33 publications

Probing Folded Proteins and Intact Protein Complexes by Desorption Electrospray Ionization Mass Spectrometry

Probing Folded Proteins and Intact Protein Complexes by Desorption Electrospray Ionization Mass Spectrometry

Mass spectrometry-enabled structural biology of membrane proteins

In situ mass spectrometry analysis of intact proteins and protein complexes from biological substrates

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