Simple Setup for Gas-Phase H/D Exchange Mass Spectrometry Coupled to Electron Transfer Dissociation and Ion Mobility for Analysis of Polypeptide Structure on a Liquid Chromatographic Time Scale

Mistarz, Ulrik H.; Brown, Jeffery M.; Haselmann, Kim F.; Rand, Kasper D.

doi:10.1021/ac5035456

Cited by 36 publications

(64 citation statements)

References 49 publications

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“…greater HDX levels do not necessarily correspond to a more extended conformer). Despite complications relating HDX levels to protein conformation, gas-phase HDX-MS has been applied to infer structural features of proteins [79].…”

Section: Gas-phase Hdx-msmentioning

confidence: 99%

Mass spectrometric methods to analyze the structural organization of macromolecular complexes

Rajabi

Ashcroft

Radford

2015

Methods

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With the development of soft ionization techniques such as electrospray ionization (ESI), mass spectrometry (MS) has found widespread application in structural biology. The ability to transfer large biomolecular complexes intact into the gas-phase, combined with the low sample consumption and high sensitivity of MS, has made ESI-MS a method of choice for the characterization of macromolecules. This paper describes the application of MS to study large non-covalent complexes. We categorize the available techniques in two groups. First, solution-based techniques in which the biomolecules are labeled in solution and subsequently characterized by MS. Three MS-based techniques are discussed, namely hydroxyl radical footprinting, cross-linking and hydrogen/deuterium exchange (HDX) MS. In the second group, MS-based techniques to probe intact biomolecules in the gas-phase, e.g. side-chain microsolvation, HDX and ion mobility spectrometry are discussed. Together, the approaches place MS as a powerful methodology for an ever growing plethora of structural applications.

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Section: Gas-phase Hdx-msmentioning

confidence: 99%

Mass spectrometric methods to analyze the structural organization of macromolecular complexes

Rajabi

Ashcroft

Radford

2015

Methods

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“…In the future, free electron lasers may provide atomically resolved images of solvent-free biomolecules [11], but at present other approaches have to be used. These include spectroscopic tools [12][13][14], dissociation experiments [5,6,8], gas phase H/D exchange [15,16], soft landing/EM [4,17], and computer simulations [10,[18][19][20]. The technique that is currently most widely used for characterizing biomolecular conformations in the gas phase is ion mobility spectrometry (IMS) [21][22][23][24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Protein Structural Studies by Traveling Wave Ion Mobility Spectrometry: A Critical Look at Electrospray Sources and Calibration Issues

Sun

Vahidi

Sowole

et al. 2015

J. Am. Soc. Mass Spectrom.

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Abstract. The question whether electrosprayed protein ions retain solution-like conformations continues to be a matter of debate. One way to address this issue involves comparisons of collision cross sections (Ω) measured by ion mobility spectrometry (IMS) with Ω values calculated for candidate structures. Many investigations in this area employ traveling wave IMS (TWIMS). It is often implied that nanoESI is more conducive for the retention of solution structure than regular ESI. Focusing on ubiquitin, cytochrome c, myoglobin, and hemoglobin, we demonstrate that Ω values and collisional unfolding profiles are virtually indistinguishable under both conditions. These findings suggest that gas-phase structures and ion internal energies are independent of the type of electrospray source. We also note that TWIMS calibration can be challenging because differences in the extent of collisional activation relative to drift tube reference data may lead to ambiguous peak assignments. It is demonstrated that this problem can be circumvented by employing collisionally heated calibrant ions. Overall, our data are consistent with the view that exposure of native proteins to electrospray conditions can generate kinetically trapped ions that retain solution-like structures on the millisecond time scale of TWIMS experiments.

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“…Indeed, using this approach researchers reported the deuterium uptake for specific ion conformers that were representative of solution conformers for ubiquitin ions. More recently, Rand and coworkers developed a rapid method for gas-phase HDX enabling the analysis of proteins and peptides distinguished by condensed-phase separations [16]. …”

Section: Introductionmentioning

confidence: 99%

Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of Anions: Part 2. Assessing Charge Site Location and Isotope Scrambling

Khakinejad

Kondalaji

Donohoe

et al. 2016

J. Am. Soc. Mass Spectrom.

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Ion mobility spectrometry (IMS) coupled with gas-phase hydrogen deuterium exchange (HDX)-mass spectrometry (MS) and molecular dynamic simulations (MDS) has been used for structural investigation of anions produced by electrospraying a sample containing a synthetic peptide having the sequence KKDDDDDIIKIIK. In these experiments the potential of the analytical method for locating charge sites on ions as well as for utilizing collision-induced dissociation (CID) to reveal the degree of deuterium uptake within specific amino acid residues has been assessed. For diffuse (i.e., more elongated) [M-2H]2− ions, decreased deuterium content along with MDS data suggest that the D4 and D6 residues are charge sites whereas for the more diffuse [M-3H]3− ions, the data suggest that the D4, D7, and the C-terminus are deprotonated. Fragmentation of mobility-selected, diffuse [M-2H]2− ions to determine deuterium uptake at individual amino acid residues reveals a degree of deuterium retention at incorporation sites. Although the diffuse [M-3H]3− ions may show more HD scrambling, it is not possible to clearly distinguish HD scrambling from the expected deuterium uptake based on a hydrogen accessibility model. The capability of the IMS-HDX-MS/MS approach to provide relevant details about ion structure is discussed. Additionally, the ability to extend the approach for locating protonation sites on positively-charged ions is presented.

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Simple Setup for Gas-Phase H/D Exchange Mass Spectrometry Coupled to Electron Transfer Dissociation and Ion Mobility for Analysis of Polypeptide Structure on a Liquid Chromatographic Time Scale

Cited by 36 publications

References 49 publications

Mass spectrometric methods to analyze the structural organization of macromolecular complexes

Mass spectrometric methods to analyze the structural organization of macromolecular complexes

Protein Structural Studies by Traveling Wave Ion Mobility Spectrometry: A Critical Look at Electrospray Sources and Calibration Issues

Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of Anions: Part 2. Assessing Charge Site Location and Isotope Scrambling

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