Coupling 193 nm Ultraviolet Photodissociation and Ion Mobility for Sequence Characterization of Conformationally-Selected Peptides

Stiving, Alyssa Q.; Harvey, Sophie R.; Jones, Benjamin J.; Bellina, Bruno; Brown, Jeffery M.; Barran, Perdita E.; Wysocki, Vicki H.

doi:10.1021/jasms.0c00259

Cited by 17 publications

(34 citation statements)

References 49 publications

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“…S7 †) based on production and consideration of both apo sequence ions (no Zn) and holo sequence ions (Zn retained). In agreement with previous studies, [74][75][76] we observed little change in UVPD sequence coverage based on precursor charge state (Fig. S8a and b †).…”

Section: Resultssupporting

confidence: 93%

Characterization of the T4 gp32–ssDNA complex by native, cross-linking, and ultraviolet photodissociation mass spectrometry

et al. 2021

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Section: Resultssupporting

confidence: 93%

Characterization of the T4 gp32–ssDNA complex by native, cross-linking, and ultraviolet photodissociation mass spectrometry

et al. 2021

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“…In most prior applications, UVPD is carried out at comparatively high vacuum of the HCD or C‐trap of an Orbitrap instrument or in an ICR cell. Even when coupling UVPD with ion mobility spectrometry, the UV irradiation event is most often carried out separately from the ion mobility device in a trap that operates at much lower pressure relative to the ion mobility separation 30–32,34–37 . The pressure conditions likely significantly impact the photoactivation of ions.…”

Section: Resultsmentioning

confidence: 99%

“…UVPD has previously been coupled with ion mobility spectrometry/mass spectrometry (IMS/MS) 29–37 . IMS/MS determines the momentum transfer cross section of an ion from measuring the velocity it acquires when traversing a gas‐filled chamber under the influence of an electric field 38 .…”

Section: Introductionmentioning

confidence: 99%

“…Various implementations have been reported; for example, UVPD was performed in quadrupolar ion traps/guides attached to the exit of drift tubes. 34,35 Alternatively, UVPD was coupled with traveling wave IMS/MS instrumentation; here, UVPD was carried out in both the pre-IMS "trap" cell 36,37 and the post-IMS "transfer" cell [30][31][32] of the "triwave" of a Synapt G2. The benefit of performing ion mobility separation upstream of UVPD is that this enables conformationally selective UVPD experiments.…”

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confidence: 99%

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Tandem‐trapped ion mobility spectrometry/mass spectrometry coupled with ultraviolet photodissociation

Liu

Ridgeway

Winfred

et al. 2021

Rapid Comm Mass Spectrometry

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Rationale Tandem‐ion mobility spectrometry/mass spectrometry methods have recently gained traction for the structural characterization of proteins and protein complexes. However, ion activation techniques currently coupled with tandem‐ion mobility spectrometry/mass spectrometry methods are limited in their ability to characterize structures of proteins and protein complexes. Methods Here, we describe the coupling of the separation capabilities of tandem‐trapped ion mobility spectrometry/mass spectrometry (tTIMS/MS) with the dissociation capabilities of ultraviolet photodissociation (UVPD) for protein structure analysis. Results We establish the feasibility of dissociating intact proteins by UV irradiation at 213 nm between the two TIMS devices in tTIMS/MS and at pressure conditions compatible with ion mobility spectrometry (2–3 mbar). We validate that the fragments produced by UVPD under these conditions result from a radical‐based mechanism in accordance with prior literature on UVPD. The data suggest stabilization of fragment ions produced from UVPD by collisional cooling due to the elevated pressures used here (“UVnoD2”), which otherwise do not survive to detection. The data account for a sequence coverage for the protein ubiquitin comparable to recent reports, demonstrating the analytical utility of our instrument in mobility‐separating fragment ions produced from UVPD. Conclusions The data demonstrate that UVPD carried out at elevated pressures of 2–3 mbar yields extensive fragment ions rich in information about the protein and that their exhaustive analysis requires IMS separation post‐UVPD. Therefore, because UVPD and tTIMS/MS each have been shown to be valuable techniques on their own merit in proteomics, our contribution here underscores the potential of combining tTIMS/MS with UVPD for structural proteomics.

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“…Using TIMS, we observed baseline IMS resolution for desR1 and desR9 bradykinin, previously unachieved on a TWIMS platform. 47 Despite having the same nominal mass, desArg1 and desArg9 yield significantly different dissociation spectra. Most notable is the preferential formation of y-type ions from desArg1 because the charge is sequestered on the C-terminal arginine.…”

Section: Tims-sid Of Bradykinin Desarg1 and Desarg9mentioning

confidence: 99%

Surface-Induced Dissociation of Protein Complexes Selected by Trapped Ion Mobility Spectrometry

Panczyk¹,

Snyder²,

Ridgeway³

et al. 2020

Preprint

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<p><a>Native mass spectrometry, particularly in conjunction with gas-phase ion mobility spectrometry measurements, has proven useful as a structural biology tool for evaluating the stoichiometry, conformation, and topology of protein complexes. Here, we demonstrate the combination of trapped ion mobility spectrometry (TIMS) and surface-induced dissociation (SID) on a Bruker SolariX XR 15 T FT-ICR mass spectrometer for structural analysis of protein complexes. We successfully performed SID on mobility-selected protein complexes, including streptavidin tetramer and cholera toxin B with bound ligand. Additionally, TIMS-SID was employed on a mixture of peptides bradykinin desR1 and desR9 to mobility separate and identify the individual peptides. Importantly, results show that native-like conformations can be maintained throughout the TIMS analysis. The TIMS-SID spectra are analogous to SID spectra acquired using quadrupole mass selection, indicating little measurable, if any, structural rearrangement during mobility selection. Mobility parking was used on the ion or mobility of interest and 50 to 200 SID mass spectra were averaged. High quality TIMS-SID spectra were acquired over a period of 2-10 minutes, comparable to or slightly longer than SID coupled with ion mobility on various instrument platforms in our laboratory. The ultrahigh resolving power of the 15 T FT-ICR allowed for the identification and relative quantification of overlapping SID fragments with the same nominal <i>m/z</i> based on isotope patterns and shows promise as a platform to probe small mass differences, such as protein-ligand binding or post-translational modifications. These results represent the potential of TIMS-SID-MS for the analysis of both protein complexes and peptides.</a></p>

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Coupling 193 nm Ultraviolet Photodissociation and Ion Mobility for Sequence Characterization of Conformationally-Selected Peptides

Cited by 17 publications

References 49 publications

Characterization of the T4 gp32–ssDNA complex by native, cross-linking, and ultraviolet photodissociation mass spectrometry

Characterization of the T4 gp32–ssDNA complex by native, cross-linking, and ultraviolet photodissociation mass spectrometry

Tandem‐trapped ion mobility spectrometry/mass spectrometry coupled with ultraviolet photodissociation

Surface-Induced Dissociation of Protein Complexes Selected by Trapped Ion Mobility Spectrometry

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