Surface-induced dissociation of protein complexes on a cyclic ion mobility spectrometer

Snyder, Dalton T.; Jones, Benjamin J.; Lin, Y.; Cooper-Shepherd, Dale A.; Hewitt, Darren; Wildgoose, Jason; Brown, Jeffery M.; Langridge, James; Wysocki, Vicki H.

doi:10.1039/d1an01407b

Cited by 19 publications

(24 citation statements)

References 60 publications

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“…Surface-induced dissociation of 91 kDa homo-11mer holo TRAP 16+ (charge reduced with TEAA and with 14 equiv of trp ) on a cyclic ion mobility spectrometer yields all possible subcomplexes, requiring separation by ion mobility. Adapted with permission from ref . Copyright 2021 the Royal Society of Chemistry.…”

Section: Sid Instrumentation For Native Mass Spectrometrymentioning

confidence: 99%

Surface-induced Dissociation Mass Spectrometry as a Structural Biology Tool

2021

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Native mass spectrometry (nMS) is evolving into a workhorse for structural biology. The plethora of online and offline preparation, separation, and purification methods as well as numerous ionization techniques combined with powerful new hybrid ion mobility and mass spectrometry systems has illustrated the great potential of nMS for structural biology. Fundamental to the progression of nMS has been the development of novel activation methods for dissociating proteins and protein complexes to deduce primary, secondary, tertiary, and quaternary structure through the combined use of multiple MS/MS technologies. This review highlights the key features and advantages of surface collisions (surface-induced dissociation, SID) for probing the connectivity of subunits within protein and nucleoprotein complexes and, in particular, for solving protein structure in conjunction with complementary techniques such as cryo-EM and computational modeling. Several case studies highlight the significant role SID, and more generally nMS, will play in structural elucidation of biological assemblies in the future as the technology becomes more widely adopted. Cases are presented where SID agrees with solved crystal or cryoEM structures or provides connectivity maps that are otherwise inaccessible by “gold standard” structural biology techniques.

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Section: Sid Instrumentation For Native Mass Spectrometrymentioning

confidence: 99%

Surface-induced Dissociation Mass Spectrometry as a Structural Biology Tool

2021

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“…39 IM separates ionized oligomers of different sizes, shapes, and charges in the gas phase as they travel through the ion mobility device at different rates, and their arrival times are measured. 40 The ions then are analyzed by a mass analyzer to determine their massto-charge ratio, m/z (Figure 3A). Trimer 2AT-L and fluorescently labeled 2AT-L analogues show a charge state distribution in the mass spectrum, leading to many overlapping m/z peaks that arise from multiple assemblies of trimers.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…In native IM-MS, oligomers and noncovalent assemblies are ionized and separated without dissociation . IM separates ionized oligomers of different sizes, shapes, and charges in the gas phase as they travel through the ion mobility device at different rates, and their arrival times are measured . The ions then are analyzed by a mass analyzer to determine their mass-to-charge ratio, m / z (Figure A).…”

Section: Resultsmentioning

confidence: 99%

Elucidating the Oligomerization and Cellular Interactions of a Trimer Derived from Aβ through Fluorescence and Mass Spectrometric Studies

Guaglianone

Torrado

Lin

et al. 2022

ACS Chem. Neurosci.

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Aβ oligomers play a central role in the neurodegeneration observed with Alzheimer's disease. Our laboratory has developed covalently stabilized trimers derived from residues 17−36 of Aβ as model systems for studying Aβ oligomers. In the current study, we apply the emerging techniques of fluorescence lifetime imaging microscopy (FLIM) and native mass spectrometry (native MS) to better understand the assembly and interactions of the oligomer model system 2AT-L in aqueous solutions and with cells. 2AT-L and fluorescently labeled 2AT-L analogues assemble in the membrane-like environment of SDS-PAGE, showing diffuse bands of oligomers in equilibrium. Native ion mobility-mass spectrometry (native IM-MS) of 2AT-L allows for the identification of discrete oligomers in solution and shows similar patterns of oligomer formation between 2AT-L and fluorescently labeled analogues.Fluorescence microscopy with SH-SY5Y cells reveals that fluorescently labeled 2AT-L analogues colocalize within lysosomes. FLIM studies with phasor analysis further elucidate the assembly of 2AT-L within cells and establish the occurrence of FRET, indicating the presence of oligomers within cells. Collectively, these multiple complementary techniques help better understand the complex behavior of the 2AT-L model system.

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“…Moreover, the protein adsorbed to the material can change the material surface energy and thus prompt cell adhesion. 51 , 52 …”

Section: Discussionmentioning

confidence: 99%

Selective Polyetheretherketone Implants Combined with Graphene Cause Definitive Cell Adhesion and Osteogenic Differentiation

Su¹,

Zhang²,

Tan³

et al. 2022

IJN

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Introduction Polyetheretherketone (PEEK) has good biosafety and chemical stability for bone repair. However, PEEK is biologically inert and cannot promote bone apposition. This study investigated whether graphene-modified PEEK (G-PEEK) could improve cell adhesion and osteogenic differentiation. Methods G-PEEK was prepared by melted blending and was characterized. In vitro, the biocompatibility of G-PPEK and the ability to promote cell adhesion and osteogenic differentiation in rabbit bone marrow mesenchymal stem cells (rBMSCs) were examined using live and dead cell double staining, the cell counting kit-8 (CCK-8) assay, immunofluorescence and quantitative real-time PCR (qRT‒PCR). An in vivo rabbit extra-articular graft-to-bone healing model was established. At 4 and 12 weeks after surgery, CT analysis and histological evaluation were performed. Results In vitro, G-PEEK significantly improved the adhesion and proliferation of rBMSCs, with good biocompatibility. In vivo, G-PEEK promoted new bone formation at the site of the bone defect. Conclusion G-PEEK showed excellent osteogenesis performance, which promises new applications in implant materials.

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Surface-induced dissociation of protein complexes on a cyclic ion mobility spectrometer

Abstract: Characterization of protein assemblies and amyloid aggregates by CIU, CID, SIU, and SID on a cyclic ion mobility spectrometer.

Cited by 19 publications

References 60 publications

Surface-induced Dissociation Mass Spectrometry as a Structural Biology Tool

Surface-induced Dissociation Mass Spectrometry as a Structural Biology Tool

Elucidating the Oligomerization and Cellular Interactions of a Trimer Derived from Aβ through Fluorescence and Mass Spectrometric Studies

Selective Polyetheretherketone Implants Combined with Graphene Cause Definitive Cell Adhesion and Osteogenic Differentiation

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