Ion Mobility Measurements and their Applications to Clusters and Biomolecules

Clemmer, David E.; Jarrold, Martin F.

doi:10.1002/(sici)1096-9888(199706)32:6<577::aid-jms530>3.3.co;2-w

Cited by 316 publications

(518 citation statements)

References 11 publications

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“…IMS coupled with ESI-MS (ESI-IMS-MS) is a powerful technique for analyzing the conformation of a protein (Knapman et al, 2010;Smith, Giles, Bateman, Radford, & Ashcroft, 2007). IMS separates ions traveling through an electric field in a drift tube according to differences in their shapes and charges (Clemmer & Jarrold, 1997).…”

Section: Retention and Collision Cross Section Of Glycopeptidesmentioning

confidence: 99%

Retention of glycopeptides analyzed using hydrophilic interaction chromatography is influenced by charge and carbon chain length of ion‐pairing reagent for mobile phase

Furuki

Toyo’oka

2017

Biomedical Chromatography

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Characterization of the glycans of glycoproteins is essential for the development and production of biologics. Numerous methods are available for analyzing the glycans of glycoproteins directly and labeled glycans. Nevertheless, glycopeptides are difficult to resolve because of their exceptional complexity and the microheterogeneity of glycans. These properties represent technical challenges to efforts to insure the accurate characterization of biopharmaceuticals to comply with regulatory requirements. Therefore, we investigated the retention behavior of peptides and glycopeptides in hydrophilic interaction chromatography-mode HPLC in the presence of ion-pairing reagents. Anionic ion-pairing reagents decreased the retention times of glycopeptides and improved resolution in the presence of higher concentrations or hydrophobicities of ion-pairing reagent. Anionic ion-pairing reagents increased retention times of larger glycans because of their increased hydrophilicity. In contrast, in the presence of cationic ion-pairing reagents, the retention times of glycopeptides with greater numbers of sialic acid residues decreased. It is appropriate to add an anionic ion-pairing reagent to the mobile phase for good separation of glycopeptides. The collision cross-sectional area values of glycopeptides determined using electrospray ionization-ion mobility spectrometry-mass spectrometry correlated with retention times. These findings support the implementation of hydrophilic interaction chromatography-mode HPLC to improve the characterization of glycosylated biopharmaceuticals.

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Section: Retention and Collision Cross Section Of Glycopeptidesmentioning

confidence: 99%

Retention of glycopeptides analyzed using hydrophilic interaction chromatography is influenced by charge and carbon chain length of ion‐pairing reagent for mobile phase

Furuki

Toyo’oka

2017

Biomedical Chromatography

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“…Overall, these results correlate with the charge state distribution: the increased population of species C at higher charge states reflects the higher exposure of protonated residues in conformations with larger CCSs. [9,67]…”

Section: Reveal Large Differences Between Species (Approximately 7% Bmentioning

confidence: 99%

“…Hence, coupled IMMS devices allow the simultaneous separation of gaseous protein ions not only on the basis of their mass but also on their size and shape. [8] IMMS has been successfully applied in the structural characterization of proteins and non-covalent protein complexes, [9][10][11] in the study of the self-aggregation of pathogenic proteins, [12][13][14] and in the detection of non-canonical DNA secondary structures, [15,16] among others. [17] Conformational dynamics is crucial for the biological function of proteins.…”

Section: Introductionmentioning

confidence: 99%

Analyzing slowly exchanging protein conformations by ion mobility mass spectrometry: study of the dynamic equilibrium of prolyl oligopeptidase

et al. 2016

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Ion mobility mass spectrometry (IMMS) is a biophysical technique that allows the separation of isobaric species on the basis of their size and shape. The high separation capacity, sensitivity and relatively fast time scale measurements confer IMMS great potential for the study of proteins in slow (µs-ms) conformational equilibrium in solution. However, the use of this technique for examining dynamic proteins is still not generalized. One of the major limitations is the instability of protein ions in the gas phase, which raises the question as to what extent the structures detected reflect those in solution. Here, we addressed this issue by analyzing the conformational landscape of prolyl oligopeptidase (POP) - a model of a large dynamic enzyme in the µs-ms range - by native IMMS and compared the results obtained in the gas phase with those obtained in solution. In order to interpret the experimental results, we used theoretical simulations. In addition, the stability of POP gaseous ions was explored by charge reduction and collision-induced unfolding experiments. Our experiments disclosed two species of POP in the gas phase, which correlated well with the open and closed conformations in equilibrium in solution; moreover, a gas-phase collapsed form of POP was also detected. Therefore, our findings not only support the potential of IMMS for the study of multiple co-existing conformations of large proteins in slow dynamic equilibrium in solution but also stress the need for careful data analysis to avoid artifacts. Copyright © 2016 John Wiley & Sons, Ltd.

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“…It is now well established that MS can yield insights into the composition, stoichiometry and connectivity of heterogeneous multiprotein assemblies at relatively low concentrations (Aquilina et al 2003; Heck and van den Heuvel 2004; Hernandez et al 2006; Sharon and Robinson 2007; Zhou et al 2008; Hernandez et al 2009). When combined with IM, it becomes possible to separate species not only according to their mass-to-charge ratio (m/z) but also according to their ability to traverse an ion guide containing inert gas under the influence of a weak electric field, thus yielding ion size and shape information (Clemmer and Jarrold 1997; Wyttenbach and Bowers 2007; Kaddis et al 2007; Bohrer et al 2008; Scarff et al 2009; Ekeowa et al 2010; Uetrecht et al 2010; Knapman et al 2010; Smith et al 2010; Cole et al 2010). Traveling wave IM separations specifically, that utilize a series of low-voltage ‘waves’ to propel ions for such size-dependant separations, have enabled most of the modern applications of IM-MS to structural biology (Giles et al 2004; Scarff et al 2008; Zhong et al 2011; Giles et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Traveling-wave ion mobility-mass spectrometry reveals additional mechanistic details in the stabilization of protein complex ions through tuned salt additives

Han

Ruotolo

2013

Int. J. Ion Mobil. Spec.

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Ion mobility–mass spectrometry is often applied to the structural elucidation of multiprotein assemblies in cases where X-ray crystallography or NMR experiments have proved challenging. Such applications are growing steadily as we continue to probe regions of the proteome that are less-accessible to such high-resolution structural biology tools. Since ion mobility measures protein structure in the absence of bulk solvent, strategies designed to more-broadly stabilize native-like protein structures in the gas-phase would greatly enable the application of such measurements to challenging structural targets. Recently, we have begun investigating the ability of salt-based solution additives that remain bound to protein ions in the gas-phase to stabilize native-like protein structures. These experiments, which utilize collision induced unfolding and collision induced dissociation in a tandem mass spectrometry mode to measure protein stability, seek to develop a rank-order similar to the Hofmeister series that categorizes the general ability of different anions and cations to stabilize gas-phase protein structure. Here, we study magnesium chloride as a potential stabilizing additive for protein structures in vacuo, and find that the addition of this salt to solutions prior to nano-electrospray ionization dramatically enhances multiprotein complex structural stability in the gas-phase. Based on these experiments, we also refine the physical mechanism of cation-based protein complex ion stabilization by tracking the unfolding transitions experienced by cation-bound complexes. Upon comparison with unbound proteins, we find strong evidence that stabilizing cations act to tether protein complex structure. We conclude by putting the results reported here in context, and by projecting the future applications of this method.

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Ion Mobility Measurements and their Applications to Clusters and Biomolecules

Cited by 316 publications

References 11 publications

Retention of glycopeptides analyzed using hydrophilic interaction chromatography is influenced by charge and carbon chain length of ion‐pairing reagent for mobile phase

Retention of glycopeptides analyzed using hydrophilic interaction chromatography is influenced by charge and carbon chain length of ion‐pairing reagent for mobile phase

Analyzing slowly exchanging protein conformations by ion mobility mass spectrometry: study of the dynamic equilibrium of prolyl oligopeptidase

Traveling-wave ion mobility-mass spectrometry reveals additional mechanistic details in the stabilization of protein complex ions through tuned salt additives

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