Applications of Charge Detection Mass Spectrometry in Molecular Biology and Biotechnology

Jarrold, Martin F.

doi:10.1021/acs.chemrev.1c00377

Cited by 80 publications

(97 citation statements)

References 267 publications

(482 reference statements)

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“…Electrospray ionization (ESI) is renowned for its ability to transfer intact proteins and protein complexes from the solution to the gas phase . Multiple charging is beneficial in tandem mass spectrometry (MS/MS) measurements for enhancing sequence information in charge-state-dependent fragmentation methods ( e.g ., electron capture dissociation, electron transfer dissociation, and ultraviolet photodissociation at lower pressures), − and signal increases proportionally with charge state for charge sensitive detection ( e.g ., FT-ICR, Orbitrap and charge detection MS). Protein ions formed from denaturing solutions typically have higher and broader charge state distributions than those formed from native-like solutions because the unfolded conformations have higher surface areas and thus, more protonation sites can be accommodated .…”

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confidence: 99%

On the Mechanism of Theta Capillary Nanoelectrospray Ionization for the Formation of Highly Charged Protein Ions Directly from Native Solutions

et al. 2022

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Theta capillary nanoelectrospray ionization (θ-nanoESI) can be used to “supercharge” protein ions directly from solution for detection by mass spectrometry (MS). In native top-down MS, the extent of protein charging is low. Given that ions with more charge fragment more readily, increasing charge can enhance the extent of sequence information obtained by top-down MS. For θ-nanoESI, dual-channeled nanoESI emitters are used to mix two solutions in low to sub-μs prior to MS. The mechanism for θ-nanoESI mixing has been reported to primarily occur: (i) in a single shared Taylor cone and in the droplets formed from the Taylor cone or (ii) by the fusion of droplets formed from two separate Taylor cones. Using θ-nanoESI-ion mobility MS, native protein solutions were rapidly mixed with denaturing supercharging solutions to form protein ions in significantly higher charge states and with more elongated structures than those formed by premixing the solutions prior to nanoESI-MS. If θ-nanoESI mixing occurred in the Taylor cone and in the droplets resulting from the single Taylor cone, then the extent of protein charging and unfolding should be comparable to or less than that obtained by premixing solutions. Thus, these data are consistent with mixing occurring via droplet fusion rather than in the Taylor cone prior to ESI droplet formation. These data also suggest that highly charged protein ions can be formed by the near-complete mixing of each solution. The presence of supercharging additives in premixed solutions can suppress volatile electrolyte evaporation, limiting the extent of protein charging compared to when the additive is delivered via one channel of a θ-nanoESI emitter. In θ-nanoESI, the formation of two Taylor cones can presumably result in substantial electrolyte evaporation from the ESI droplets containing native-like proteins prior to droplet fusion, thereby enhancing ion charging.

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confidence: 99%

On the Mechanism of Theta Capillary Nanoelectrospray Ionization for the Formation of Highly Charged Protein Ions Directly from Native Solutions

et al. 2022

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“…The oligomeric states of ectopically expressed portal proteins can range from 11-mers to 14-mers [ 20 , 21 , 23 , 34 , 51 ]. However, in bacteriophage P22, charge detection mass spectrometry (CDMS), which measures the mass of native protein complexes [ 44 , 45 , 46 ] was previously used to demonstrate that in vitro assembled WT portal protein rings primarily exist as 12-mers [ 15 ]. To determine if there were mutant-specific defects in portal protein polymerization, we again used CDMS to probe the oligomeric state of in vitro assembled mutant PC portal rings.…”

Section: Resultsmentioning

confidence: 99%

“…Charge detection mass spectrometry is a single particle technique where the mass-to-charge ratio ( m/z ) and charge ( z ) are simultaneously measured for each ion [ 44 , 45 , 46 ]. The m / z and z are then multiplied to give the mass of the ion.…”

Section: Methodsmentioning

confidence: 99%

Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22

Woodbury

Motwani

Leroux

et al. 2022

Viruses

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The oligomerization and incorporation of the bacteriophage P22 portal protein complex into procapsids (PCs) depends upon an interaction with scaffolding protein, but the region of the portal protein that interacts with scaffolding protein has not been defined. In herpes simplex virus 1 (HSV-1), conserved tryptophan residues located in the wing domain are required for portal-scaffolding protein interactions. In this study, tryptophan residues (W) present at positions 41, 44, 207 and 211 within the wing domain of the bacteriophage P22 portal protein were mutated to both conserved and non-conserved amino acids. Substitutions at each of these positions were shown to impair portal function in vivo, resulting in a lethal phenotype by complementation. The alanine substitutions caused the most severe defects and were thus further characterized. An analysis of infected cell lysates for the W to A mutants revealed that all the portal protein variants except W211A, which has a temperature-sensitive incorporation defect, were successfully recruited into procapsids. By charge detection mass spectrometry, all W to A mutant portal proteins were shown to form stable dodecameric rings except the variant W41A, which dissociated readily to monomers. Together, these results suggest that for P22 conserved tryptophan, residues in the wing domain of the portal protein play key roles in portal protein oligomerization and incorporation into procapsids, ultimately affecting the functionality of the portal protein at specific stages of virus assembly.

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“…1 Electrospray ionization (ESI) is renowned for its ability to transfer intact proteins and protein complexes from the solution to the gas phase. 2 Multiple charging is beneficial in tandem mass spectrometry (MS/MS) measurements for enhancing sequence information because ions in higher charge states tend to fragment more readily than those in lower charge states, [3][4][5] and signal increases proportionally with charge state for charge sensitive detection (e.g., FT-ICR, Orbitrap and charge detection MS 6 ). Protein ions formed from denaturing solutions typically have higher and broader charge state distributions than those formed from native-like solutions because the unfolded conformations have higher surface areas and thus, more protonation sites can be accommodated.…”

mentioning

confidence: 99%

On the mechanism of theta capillary nanoelectrospray ionization for the formation of highly charged protein ions directly from native solutions

Brown

Zenaidee

Loo

et al. 2022

Preprint

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Theta capillary nanoelectrospray ionization (θ-nanoESI) can be used to ‘supercharge’ protein ions directly from solution for detection by mass spectrometry (MS). In native top-down MS, the extent of protein charging is low. Given that ions with more charge fragment more readily, increasing charge can enhance the extent of sequence information obtained by top-down MS. For θ-nanoESI, dual-channelled nanoESI emitters are used to mix two solutions in low to sub-μs prior to MS. The mechanism for θ-nanoESI mixing has been reported to occur in the Taylor cone prior to ESI-droplet formation, or by the fusion of droplets formed from separate Taylor cones. Using θ-nanoESI-ion mobility-MS, native protein solutions were rapidly mixed with denaturing supercharging solutions to form protein ions in significantly higher charge states and with more elongated structures than those formed by pre-mixing the solutions prior to nanoESI-MS. If θ-nanoESI mixing occurred in the Taylor cone, then the extent of protein charging and unfolding should be comparable or less than that obtained by pre-mixing solutions. Thus, these data are consistent with mixing occurring via droplet fusion rather than in the Taylor cone prior to ESI droplet formation. The presence of supercharging additives in pre-mixed solutions can suppress volatile electrolyte evaporation, limiting the extent of protein charging compared to when the additive is delivered via one channel of a θ-nanoESI emitter. In θ-nanoESI, the formation of two Taylor cones can presumably result in substantial electrolyte evaporation from the ESI droplets containing native-like proteins prior to droplet fusion, thereby enhancing ion charging.

show abstract

Applications of Charge Detection Mass Spectrometry in Molecular Biology and Biotechnology

Cited by 80 publications

References 267 publications

On the Mechanism of Theta Capillary Nanoelectrospray Ionization for the Formation of Highly Charged Protein Ions Directly from Native Solutions

On the Mechanism of Theta Capillary Nanoelectrospray Ionization for the Formation of Highly Charged Protein Ions Directly from Native Solutions

Tryptophan Residues Are Critical for Portal Protein Assembly and Incorporation in Bacteriophage P22

On the mechanism of theta capillary nanoelectrospray ionization for the formation of highly charged protein ions directly from native solutions

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