Surface-Induced Dissociation of Anionic vs Cationic Native-Like Protein Complexes

2021 ◽  
Author(s):  
Sophie R. Harvey ◽  
Zachary L. VanAernum ◽  
Vicki H. Wysocki
Keyword(s):  
Protein Complexes ◽  
Surface Induced Dissociation

Extended Gas-Phase Trapping Followed by Surface-Induced Dissociation of Noncovalent Protein Complexes

2015 ◽  
Vol 88 (2) ◽  
pp. 1218-1221 ◽  
Author(s):  
Sophie R. Harvey ◽  
Jing Yan ◽  
Jeffery M. Brown ◽  
Emmy Hoyes ◽  
Vicki H. Wysocki
Keyword(s):  
Gas Phase ◽  
Protein Complexes ◽  
Surface Induced Dissociation ◽  
Noncovalent Protein

scholarly journals Surface-Induced Dissociation of Anionic vs Cationic Native-like Protein Complexes

2021 ◽  
Author(s):  
Sophie Harvey ◽  
Zachary VanAernum ◽  
Vicki Wysocki
Keyword(s):  
Protein Interactions ◽  
Structural Information ◽  
Protein Complexes ◽  
Charge Reduction ◽  
Surface Induced Dissociation ◽  
Negative Mode ◽  
Peak Broadening ◽  
Lower Charge ◽  
Positive Mode ◽  
Charge States

<p>Characterizing protein-protein interactions, stoichiometries, and subunit connectivity is key to understanding how subunits assemble in biologically relevant multi-subunit protein complexes. Native mass spectrometry (nMS) has emerged as a powerful tool to study protein complexes due to its low sample requirements and tolerance for heterogeneity. For such nMS studies, positive mode ionization is routinely used and charge reduction, through the addition of solution additives, is often used, as the resulting lower charge states are often more compact and considered more native like. When studied with surface-induced dissociation, charge reduced complexes often give increased structural information over their “normal-charged” counter parts. A disadvantage of charge-reduction is that increased adduction, and hence peak broadening, is often observed when charge-reducing solution additives are present. Recent studies have shown that protein complexes ionized using negative mode generally form in lower charge states relative to positive mode. Here we demonstrate that the lower charged protein complex anions, activated by SID in an ultrahigh mass range Orbitrap mass spectrometer, fragment in a manner consistent with their solved structure, hence providing substructural information. Negative mode ionization in ammonium acetate offers the advantage of charge reduction without the peak broadening associated with solution phase charge reduction additives and provides direct structural information, when coupled with SID. </p>

scholarly journals Surface-Induced Dissociation of Noncovalent Protein Complexes in an Extended Mass Range Orbitrap Mass Spectrometer

2019 ◽  
Vol 91 (5) ◽  
pp. 3611-3618 ◽  
Author(s):  
Zachary L. VanAernum ◽  
Joshua D. Gilbert ◽  
Mikhail E. Belov ◽  
Alexander A. Makarov ◽  
Stevan R. Horning ◽  
...  
Keyword(s):  
Mass Spectrometer ◽  
Protein Complexes ◽  
Mass Range ◽  
Surface Induced Dissociation ◽  
Orbitrap Mass Spectrometer ◽  
Noncovalent Protein

Illustration of SID-IM-SID (surface-induced dissociation-ion mobility-SID) mass spectrometry: homo and hetero model protein complexes

The Analyst ◽  
2015 ◽  
Vol 140 (20) ◽  
pp. 7012-7019 ◽  
Author(s):  
Royston S. Quintyn ◽  
Sophie R. Harvey ◽  
Vicki H. Wysocki
Keyword(s):  
Mass Spectrometry ◽  
Ion Mobility ◽  
Protein Complex ◽  
Protein Complexes ◽  
Second Stage ◽  
Surface Induced Dissociation ◽  
Model Protein ◽  
Complex Architecture

Surface collisions generate subcomplexes, which are then separated by ion mobility and dissociated into their individual subunitsviaa second stage of surface collisions to elucidate protein complex architecture and assembly.

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

2020 ◽  
Author(s):  
Erin Panczyk ◽  
Dalton Snyder ◽  
Mark. E. Ridgeway ◽  
Arpad Somogyi ◽  
Melvin A. Park ◽  
...  
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Small Mass ◽  
Resolving Power ◽  
Mass Selection ◽  
Trapped Ion ◽  
Surface Induced Dissociation ◽  
Trapped Ion Mobility Spectrometry ◽  
Ft Icr

<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>

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

2020 ◽  
Author(s):  
Erin Panczyk ◽  
Dalton Snyder ◽  
Mark. E. Ridgeway ◽  
Arpad Somogyi ◽  
Melvin A. Park ◽  
...  
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Small Mass ◽  
Resolving Power ◽  
Mass Selection ◽  
Trapped Ion ◽  
Surface Induced Dissociation ◽  
Trapped Ion Mobility Spectrometry ◽  
Ft Icr

<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>

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

2021 ◽  
Author(s):  
Erin M. Panczyk ◽  
Dalton T. Snyder ◽  
Mark E. Ridgeway ◽  
Árpád Somogyi ◽  
Melvin A. Park ◽  
...  
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Protein Complexes ◽  
Trapped Ion ◽  
Surface Induced Dissociation ◽  
Trapped Ion Mobility Spectrometry
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