Smaller Collision Cross Sections Research Articles

Potential of ion mobility mass spectrometry in cellulose ether analysis: substitution pattern of hydroxyethyl celluloses

Ion mobility mass spectrometry (ESI-tims-ToF–MS, syringe pump infusion) has been applied to glucose and oligosaccharide ethers derived from hydroxyethyl-methyl celluloses (HEMC) and hydroxyethyl celluloses (HEC) after permethylation and partial depolymerization: by hydrolysis without or with subsequent reductive amination with m-amino benzoic acid (mABA) or by reductive cleavage. As model compounds without tandem substitution methoxyethylated methylcellulose was used. Regioisomeric glucose ethers were separated according to their ion mobility, and positions of substitution could be assigned. Glucose ethers including isomers with tandem substitution showed additional signals with a smaller collision cross-section (CCS) than core-substituted isomers. Positional isomers of cellobiose ethers were only partly resolved due to too high complexity but showed a characteristic fingerprint that might allow classifying samples. Relative intensities of signals of glucose ether isomers could only be quantified in case of ABA derivatives with its fixed charge, while sodium adducts of methoxyethyl ethers showed an influence of the MeOEt position on ion yield. Results were in very good agreement with reference analysis. [M + Na]+ adducts of α- and β-anomers of glucose derivatives were separated in IM, complicating position assignment. This could be overcome by reductive cleavage of the permethylated HE(M)C yielding 1,5-anhydroglucitol-terminated oligosaccharides, showing the best resolved fingerprints of the cellobiose ethers of a particular cellulose ether. With this first application of ion mobility MS to the analysis of complex cellulose ethers, the promising potential of this additional separation dimension in mass spectrometry is demonstrated and discussed.Graphical abstract

Interconversions of Structural Isomers of [PdAu8(PPh3)8]2+ and [Au9(PPh3)8]3+ Revealed by Ion Mobility Mass Spectrometry

Collision cross sections (CCSs) of ligand-protected metal clusters were evaluated using ion mobility mass spectrometry. The targets used in this study were phosphine-protected clusters [PdAu8(PPh3)8]2+ and [Au9(PPh3)8]3+, for which the total structures have been resolved by single-crystal X-ray analysis. The arrival time distributions of [PdAu8(PPh3)8]2+ as a function of the He flow rate in a cell located just in front of a traveling wave ion mobility cell filled with N2 buffer gas demonstrated that it got converted to another structural isomer having a smaller CCS, with the increase in the nominal collision energy. A similar phenomenon was observed for [Au9(PPh3)8]3+. These results were explained by the collisional excitation and cooling with the buffer gas inducing the conversion of the packing arrangement of the ligands rather than the atomic structure of the metallic core: the ligand layer was converted from disordered to the closely packed arrangement found in a single crystal during this process. Th...

Folding of Protein Ions in the Gas Phase after Cation-to-Anion Proton-Transfer Reactions.

The structure and folding of a protein in solution depends on noncovalent interactions within the protein and those with surrounding ions and molecules. Decoupling these interactions in solution is challenging, which has hindered the development of accurate physics-based models for structure prediction. Investigations of proteins in the gas phase can be used to selectively decouple factors affecting the structures of proteins. Here, we use cation-to-anion proton-transfer reactions (CAPTR) to reduce the charge states of denatured ubiquitin ions in the gas phase, and ion mobility to probe their structures. In CAPTR, a precursor charge state is selected (P) and reacted with monoanions to generate charge-reduced product ions (C). Following each CAPTR event, denatured ubiquitin ions (13+ to 6+) yield products that rapidly isomerize to structures that have smaller collision cross sections (Ω). The Ω values of CAPTR product ions depend strongly on C and very weakly on P. Pre- and post-CAPTR activation was then used to probe the potential-energy surfaces of the precursor and product ions, respectively. Post-CAPTR activation showed that ions of different P fold differently and populate different regions of the potential-energy surface of that ion. Finally, pre-CAPTR activation showed that the structures of protein ions can be indirectly investigated using ion mobility of their CAPTR product ions, even for subtle structural differences that are not apparent from ion mobility characterization of the activated precursor ions. More generally, these results show that CAPTR strongly complements existing techniques for characterizing the structures and dynamics of biological molecules in the gas phase.

Separation and conversion dynamics of nuclear-spin isomers of gaseous methanol.

All symmetrical molecules with non-zero nuclear spin exist in nature as nuclear-spin isomers (NSIs). However, owing to the lack of experimental information, knowledge is rare about interconversions of NSIs of gaseous molecules with torsional symmetry. Here we report our separation and conversion observations on NSI-torsion-specific transition systems of gaseous methanol from a light-induced drift experiment involving partially spatial separation of the ortho and para isomers. We find that vibrationally excited molecules of the methanol spin isomer have a smaller collision cross-section than their ground-state counterparts. Interconversion of the enriched ortho isomer with the para isomer, which is generally considered improbable, has been quantitatively studied by sensitive detections of the spectral intensities. Rather counterintuitively, this reveals that the interconversion is inhibited with increasing pressure. Our results suggest that the spin conversion mechanism in methanol is via a quantum relaxation process with the quantum Zeno effect induced by molecular collisions.

Dissecting the dynamic conformations of the metamorphic protein lymphotactin.

A mass spectrometer provides an ideal laboratory to probe the structure and stability of isolated protein ions. Interrogation of each discrete mass/charge-separated species enables the determination of the intrinsic stability of a protein fold, gaining snapshots of unfolding pathways. In solution, the metamorphic protein lymphotactin (Ltn) exists in equilibrium between two distinct conformations, a monomeric (Ltn10) and a dimeric (Ltn40) fold. Here, we use electron capture dissociation (ECD) and drift tube ion mobility-mass spectrometry (DT IM-MS) to analyze both forms and use molecular dynamics (MD) to consider how the solution fold alters in a solvent-free environment. DT IM-MS reveals significant conformational flexibility for the monomer, while the dimer appears more conformationally restricted. These findings are supported by MD calculations, which reveal how salt bridges stabilize the conformers in vacuo. Following ECD experiments, a distinctive fragmentation pattern is obtained for both the monomer and dimer. Monomer fragmentation becomes more pronounced with increasing charge state especially in the disordered regions and C-terminal α-helix in the solution fold. Lower levels of fragmentation are seen in the β-sheet regions and in regions that contain salt bridges, identified by MD simulations. The lowest charge state of the dimer for which we obtain ECD data ([D+9H](9+)) exhibits extensive fragmentation with no relationship to the solution fold and has a smaller collision cross section (CCS) than charge states 10-13+, suggesting a "collapsed" encounter complex. Other charge states of the dimer, as for the monomer, are resistant to fragmentation in regions of β-sheets in the solution fold. This study provides evidence for preservation and loss of global fold and secondary structural elements, providing a tantalizing glimpse into the power of the emerging field of native top-down mass spectrometry.

Structure-drift time relationships in ion mobility mass spectrometry

Ion mobility spectrometry (IMS) separates ions while they travel through a buffer gas under the influence of an electrical field. The separation is affected by mass and charge but most particularly by shape (collision cross section). When coupled to MS, IMS-MS offers therefore a powerful tool for structural elucidation and isomer separation. Systematic studies aimed to compare and quantitate the effects of structural changes on drift time such as length and ramification of carbon chain, unsaturation, geometrical isomerism (cis/trans isomers for instance), cyclization and ring size are, however, scarce. Herein we used traveling wave ion mobility mass spectrometry (TWIM-MS) to systematically evaluate the relationship between structure and drift time. For that, a series of deprotonated carboxylic acids were used as model ions with a carboxylate “charge tag” for gas phase MS manipulation. Carboxylic acids showed a near linear correlation between the increase of carbon number and the increase of collision cross section (CCS). The number of double bonds changes slightly the CCS of unsaturated acids. No differences in drift time and no significant differences in CCS of cis- and trans-double bond of oleic and elaidic acids were observed. Cyclization considerably reduces the CCS. In cyclic carboxylic acids, the increase of double bonds and aromatization significantly reduces the CCS and the drift times. The use of a more polarizable drift gas, CO2, improved in some cases the separation, as for biomarker isomers of steranoic acids. The β-isomer (cis-decaline) has smaller CCS and therefore displayed lower drift time compared to the α-isomer (trans-decaline). Structural changes revealed by calculations were correlated with trends in drift times.

Charge-State Dependent Compaction and Dissociation of Protein Complexes: Insights from Ion Mobility and Molecular Dynamics

Collapse to compact states in the gas phase, with smaller collision cross sections than calculated for their native-like structure, has been reported previously for some protein complexes although not rationalized. Here we combine experimental and theoretical studies to investigate the gas-phase structures of four multimeric protein complexes during collisional activation. Importantly, using ion mobility-mass spectrometry (IM-MS), we find that all four macromolecular complexes retain their native-like topologies at low energy. Upon increasing the collision energy, two of the four complexes adopt a more compact state. This collapse was most noticeable for pentameric serum amyloid P (SAP) which contains a large central cavity. The extent of collapse was found to be highly correlated with charge state, with the surprising observation that the lowest charge states were those which experience the greatest degree of compaction. We compared these experimental results with in vacuo molecular dynamics (MD) simulations of SAP, during which the temperature was increased. Simulations showed that low charge states of SAP exhibited compact states, corresponding to collapse of the ring, while intermediate and high charge states unfolded to more extended structures, maintaining their ring-like topology, as observed experimentally. To simulate the collision-induced dissociation (CID) of different charge states of SAP, we used MS to measure the charge state of the ejected monomer and assigned this charge to one subunit, distributing the residual charges evenly among the remaining four subunits. Under these conditions, MD simulations captured the unfolding and ejection of a single subunit for intermediate charge states of SAP. The highest charge states recapitulated the ejection of compact monomers and dimers, which we observed in CID experiments of high charge states of SAP, accessed by supercharging. This strong correlation between theory and experiment has implications for further studies as well as for understanding the process of CID and for applications to gas-phase structural biology more generally.

CONTROL OF SIZE AND MORPHOLOGY OF NANO PARTICLES USING CO 2 LASER DURING FLAME SYNTHESIS

An attempt has been made to control size and morphology of nanoparticles generating and growing in a co-flow oxy-hydrogen flame combining CO 2 laser irradiation. The coalescence characteristic time could be controlled nearly independent of the collision characteristic time of particles by raising particle temperature rapidly with laser beam irradiation during flame synthesis. Silica aggregates generated by hydrolysis of SiCl 4 in a flame are irradiated by a high-power CO 2 laser beam, and rapidly heated up to temperatures that are high enough to enhance sintering, but lower than evaporation temperature. Sintering of aggregates is rapidly enhanced to change aggregates into more spherical particles. Since spherical particles have much smaller collision cross sections than volume-equivalent aggregates, much slower growth of particles is observed when the flame is irradiated by CO 2 laser beam and as a result, the diameter of resulting spherical particles decreases to about 60% of the size observed without CO 2 laser beam as the laser power increases. A higher precursor flow rate even resulted in the change of non-spherical particles into smaller spherical particles as CO 2 laser power increased. Light-scattering measurement with an Ar-ion laser and TEM observation through a localized thermophoretic sampling were used to confirm the above effects of CO 2 laser irradiation in a flame. Depending on the irradiation height of CO 2 laser beam in a flame, significantly different mechanisms were found. The radial distributions of scattering intensity and morphological change were also studied. The proposed method controlling the sintering characteristic time of particles using CO 2 laser irradiation in a flame seems to be promising to produce smaller and, at the same time, spherical nanoparticles even for high carrier gas flow rate.

Atmospheric pressure ion mobility spectrometry of protonated and sodiated peptides.

A number of peptides were studied with electrospray ionization--ion mobility spectrometry/mass spectrometry (ESI-IMS/MS). The ion mobility data were used to calculate the average collision cross sections of the different detected peptide ions in the nitrogen drift gas. By comparing the cross sections of related ions, structural information about the most probable location of the charge and the gas-phase ion conformations was deduced. For bradykinin and kemptide, a significant mobility difference between protonated and sodiated species (where sodium replaced a proton in singly and doubly charged peptides) was demonstrated. Surprisingly, the doubly charged sodiated peptides had a smaller collision cross section than the doubly charged protonated ones leading to the conclusion that the gas-phase conformations of these ions are different with respect to intramolecular interactions.