Accumulating evidence suggests that solution-phase conformations of small globular proteins and large molecular protein assemblies can be preserved for milliseconds after electrospray ionization. Thus, the study in gas phase on this time scale is highly desirable. Here we demonstrate a traveling wave ion guide (TWIG) Synapt mass spectrometer offers suitable environment rapid efficient gas-phase hydrogen/deuterium exchange (HDX). Gaseous ND3 was introduced into either source TWIG or located...

Top-down approaches for the characterization of intact proteins and macromolecular complexes are becoming increasingly popular, since they potentially simplify speed up assignment process. Here we demonstrate how, on a commercially available Q-TWIMS-TOF instrument, performed top-down ETD native form tetrameric alcohol dehydrogenase. We achieved good sequence coverage throughout first 81 N-terminal amino acids ADH, with exception loop located inside protein. This is in agreement exposed parts...

Non-dissociative charge reduction, typically considered to be an unwanted side reaction in electron transfer dissociation (ETD) experiments, can enhanced significantly order reduce the state of intact protein complexes as low 1+ on a commercially available Q-IM-TOF instrument. This allows for detection large beyond 100,000 m/z, while at same time generating top-down ETD fragments, which provide sequence information from surface-exposed parts folded structure. Optimization supplemental...

Electron-based fragmentation methods have revolutionized biomolecular mass spectrometry, in particular native and top-down protein analysis. Here, we report the use of a new electromagnetostatic cell to perform electron capture dissociation (ECD) within quadrupole/ion mobility/time-of-flight spectrometer. This was installed between ion mobility time-of-flight regions instrument, fast enough be compatible with separation. The instrument already fitted transfer (ETD) quadrupole prior...

The recent application of electron transfer dissociation (ETD) to measure the hydrogen exchange proteins in solution at single-residue resolution (HX-ETD) paves way for mass spectrometry-based analyses biomolecular structure an unprecedented level detail. approach requires that activation polypeptide ions prior ETD is minimal so as prevent undesirable gas-phase randomization deuterium label from (i.e., scrambling). Here we explore use a traveling wave ion guide quadrupole-time-of-flight...

Electron transfer dissociation (ETD) and collision induced (CID) have been used to locate the precise binding sites for platinum ruthenium anticancer complexes on peptide Substance P. We show that ETD combined with ion mobility-mass spectrometry significantly reduces mass spectral complexity improves S/N of product-ions formed.

To interpret the wealth of information contained in hydrogen/deuterium exchange (HDX) behavior peptides and proteins gas-phase, analytical tools are needed to resolve HDX individual exchanging sites. Here we show that ETD can be combined with fast gas-phase ND(3) gas used monitor side-chain hydrogens residues both small peptide ions larger protein a few milliseconds after electrospray. By employing consecutive traveling wave ion guides mass spectrometer, were labeled on-the-fly (0.1-10 ms)...

An ion mobility mass spectrometer has been modified to allow optical interrogation of ions with different mass-to-charge ratios and/or mobilities.

Mass spectrometry (MS) allows for automated analysis of complex samples at high resolution without the need labeling/derivatization. Liquid atmospheric pressure matrix-assisted laser desorption/ionization (LAP-MALDI) enables rapid sample preparation and MS using microtiter-plate formats high-performing mass spectrometers. We present a step change in high-speed, large-scale peptides 20 samples/s an enzymatic assay 40 samples/s, i.e., order magnitude faster than current platforms. LAP-MALDI...

Ultraviolet matrix-assisted laser desorption/ionization-mass spectrometry has been employed with time-lag focusing to explore its utility for the characterization of synthetic polymers broad distributions. Mixtures five polymer standards narrow molecular weight distributions were analyzed. The spectra found be broadly those expected three different types systems—poly(styrene), poly(methyl methacrylate), and poly(ethylene glycol)—when equimass mixtures used. Large changes in apparent...

The initial stages of protein unfolding may reflect the stability entire fold and can also reveal which parts a be perturbed, without restructuring rest. In this work, we couple UVPD with activated ion mobility mass spectrometry to measure how three model proteins start unfold. Ubiquitin, cytochrome c myoglobin ions produced via nESI from salty solutions are subjected UV irradiation pre-mobility separation; experiments conducted range source conditions alter conformation precursor as shown...

The position of C=C within fatty acyl chains affects the biological function lipids. Ozone-induced dissociation mass spectrometry (OzID-MS) has great potential in determination lipid double-bond position, but generally been implemented on low-resolution ion trap spectrometers. In addition, most OzID-MS experiments carried out so far were focused sodiated adducts lipids; fragmentation commonly observed protonated ions generated LC/MS-based lipidomics workflow less explored.

Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is now a routinely used technique to inform on protein structure, dynamics, and interactions. Localizing the incorporated deuterium content single residue basis increases spatial resolution of this enabling detailed structural analysis. Here, we investigate use ultraviolet photodissociation (UVPD) at 213 nm measure levels in HDX-MS experiments. Using selectively labeled peptide, show that UVPD occurs without H/D scrambling as peptide...

Hydrogen/deuterium exchange (HDX) mass spectrometry (MS) for protein structural analysis has been adopted many purposes, including biopharmaceutical development. One of the benefits examining amide proton by is that it can readily resolve different regimes, as evidenced either binomial or bimodal isotope patterns. By careful pattern during exchange, more insight be obtained on behavior in solution. However, one must sure any observed patterns are not artifacts and reflective true Sample...

Tandem mass spectrometry (MS/MS) is an invaluable experimental tool for providing analytical data supporting the identification of small molecules and peptides in mass-spectrometry-based "omics" experiments. Data-dependent MS/MS (DDA) a real-time MS/MS-acquisition strategy that responsive to signals detected given sample. However, analysis even moderately complex samples with state-of-the-art instrumentation, speed acquisition insufficient offer comprehensive coverage all molecules....

We demonstrate the capabilities of a laser-coupled ion mobility mass spectrometer for analysis peptide sequence and structure showing ultraviolet photodissociation (UVPD) spectra selected ions. A Synapt G2-S has been modified to allow photointeraction ions post cell. For this work, we have employed single wavelength laser, which irradiates at 266 nm. present unique instrument several key features. Irradiation luteinizing hormone releasing (LHRH), growth hexapeptide (GHRP-6), TrpCage...

Label-free high-throughput screening using mass spectrometry has the potential to provide rapid large-scale sample analysis at a speed of more than one per second. Such is important for compound library, assay and future clinical millions samples within reasonable time frame. Herein, we present liquid atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) setup (>5 second) three substance classes (peptides, antibiotics, lipids). Liquid support matrices (LSM) were used...

Infrared multiphoton dissociation (IRMPD) spectroscopy, using a free-electron laser, and ion mobility measurements, both drift-cell traveling-wave instruments, were used to investigate the structure of gas-phase peptide (AAHAL + 2H)(2+) ions produced by electrospray ionization. The experimental data from IRMPD spectra collisional cross section (Ω) measurements consistent with respective infrared Ω calculated for lowest-energy conformer obtained extensive molecular dynamics searches combined...

Near-infrared surface plasmon resonance (SPR) spectra were collected of thin multilayer films aluminum-doped zinc oxide (AZO) / silver (Ag) AZO on BK-7 glass in the Kretschmann configuration air, with layer thickness varying from 5 nm to 50 nm. The SPR results interpreted by modeling reflectance a five-layer transfer-matrix method, aid simplex algorithm. model indicated that Ag plasma frequency was significantly higher than bulk value, possibly due Schottky effect charge transfer layer....

Gas-phase hydrogen/deuterium exchange (HDX) is a fast and sensitive, yet unharnessed analytical approach for providing information on the structural properties of biomolecules, in complementary manner to mass analysis. Here, we describe simple setup ND3-mediated millisecond gas-phase HDX inside spectrometer immediately after ESI (gas-phase HDX-MS) show utility studying primary higher-order structure peptides proteins. was achieved by passing N2-gas through container filled with aqueous...