Electron capture dissociation of polypeptides using a 3 Tesla Fourier transform ion cyclotron resonance mass spectrometer

Electron capture dissociation (ECD) of polypeptides has been demonstrated using a commercially available 3 Tesla Fourier transform ion cyclotron resonance (FTICR) instrument. A conventional rhenium filament, designed for high-energy electron impact ionisation, was used to effect ECD of substance P, bee venom melittin and bovine insulin, oxidised B chain. A retarding field analysis of the effective electron kinetic energy distribution entering the ICR cell suggests that one of the most important parameters governing ECD for this particular instrument is the need to employ low trapping plate voltages. This is shown to maximise the abundance of low-energy electrons. The demonstration of ECD at this relatively low magnetic field strength could offer the prospect of more routine ECD analysis for the wider research community, given the reduced cost of such magnets and (at least theoretically) the greater ease of electron/ion cloud overlap at lower field.     

IRMPD Action Spectroscopy of Alkali Metal Cation–Cytosine Complexes: Effects of Alkali Metal Cation Size on Gas Phase Conformation

The gas-phase structures of alkali metal cation-cytosine complexes generated by electrospray ionization are probed via infrared multiple photon dissociation (IRMPD) action spectroscopy and theoretical calculations. IRMPD action spectra of five alkali metal cation–cytosine complexes exhibit both similar and distinctive spectral features over the range of ~1000–1900 cm^-1. The IRMPD spectra of the Li⁺(cytosine), Na⁺(cytosine), and K⁺(cytosine) complexes are relatively simple but exhibit changes in the shape and shifts in the positions of several bands that correlate with the size of the alkali metal cation. The IRMPD spectra of the Rb⁺(cytosine) and Cs⁺(cytosine) complexes are much richer as distinctive new IR bands are observed, and the positions of several bands continue to shift in relation to the size of the metal cation. The measured IRMPD spectra are compared to linear IR spectra of stable low-energy tautomeric conformations calculated at the B3LYP/def2-TZVPPD level of theory to identify the conformations accessed in the experiments. These comparisons suggest that the evolution in the features in the IRMPD action spectra with the size of the metal cation, and the appearance of new bands for the larger metal cations, are the result of the variations in the intensities at which these complexes can be generated and the strength of the alkali metal cation-cytosine binding interaction, not the presence of multiple tautomeric conformations. Only a single tautomeric conformation is accessed for all five alkali metal cation–cytosine complexes, where the alkali metal cation binds to the O2 and N3 atoms of the canonical amino-oxo tautomer of cytosine, M⁺(C₁).

Inert site in a protein zinc cluster: isotope exchange by high resolution mass spectrometry

Using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry, we show that one Zn2+ ion in the Zn4 cluster of cyanobacterial metallothionein is inert to exchange with 67Zn2+. We suggest that this is zinc in site A, which together with the surrounding alpha and beta secondary structure forms a zinc-finger fold.     

A new ternary Cu (II) complex with 4,7-dimethyl-1,10-phenanthroline and NOO-type tridentate Schiff base ligand: Synthesis,crystal structure,biomacromolecular interactions,and radical scavenging activities

NOO-type tridentate Schiff base, N-salicylidene-2-aminobenzoic acid, (H₂L), and its ternary Cu (II) complex containing H₂L Schiff base and 4,7-dimethyl-1,10-phenanthroline (4,7-dmphen), [Cu(4,7-dmphen)(H₂L)]₂7H₂O, have been synthesized and characterized by CHN analysis, ESI-MS, FTIR, and single-crystal X-ray diffraction techniques. The interaction of alone H₂L Schiff base ligand and ternary Cu (II) complex with biomacramolecules {calf thymus DNA (CT-DNA) and bovine serum albumin (BSA)} has been investigated by electronic absorption and fluorescence spectroscopy. The experimental results indicate that H₂L Schiff base ligand and ternary Cu (II) complex bind to CT-DNA by means of a moderate intercalation mode. Furthermore, the fluorescence quenching mechanism between H₂L Schiff base ligand and ternary Cu (II) complex with BSA possesses a static quenching process. Radical scavenging activity of H₂L Schiff base ligand and ternary Cu (II) complex was measured in terms of EC₅₀, using the DPPH and H₂O₂ methods. Biomacromolecule interactions and scavenging activity studies revealed that ternary Cu (II) complex yielded better results than H₂L Schiff base ligand alone.     

Radical Rearrangement Chemistry in Ultraviolet Photodissociation of Iodotyrosine Systems: Insights from Metastable Dissociation,Infrared Ion Spectroscopy,and Reaction Pathway Calculations

We report on the ultraviolet photodissociation (UVPD) chemistry of protonated tyrosine, iodotyrosine, and diiodotyrosine. Distonic loss of the iodine creates a high-energy radical at the aromatic ring that engages in hydrogen/proton rearrangement chemistry. Based on UVPD kinetics measurements, the appearance of this radical is coincident with the UV irradiation pulse (8 ns). Conversely, sequential UVPD product ions exhibit metastable decay on ca. 100 ns timescales. Infrared ion spectroscopy is capable of confirming putative structures of the rearrangement products as proton transfers from the imine and β-carbon hydrogens. Potential energy surfaces for the various reaction pathways indicate that the rearrangement chemistry is highly complex, compatible with a cascade of rearrangements, and that there is no preferred rearrangement pathway even in small molecular systems like these.

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Mid-infrared vibrational spectra of discrete acetone-ligated cerium hydroxide cations

Cerium(iii) hydroxy reactive sites are responsible for several important heterogeneous catalysis processes, and understanding the reaction chemistry of substrate molecules like CO, H(2)O, and CH(3)OH as they occur in heterogeneous media is a challenging task. We report here the first infrared spectra of model gas-phase cerium complexes and use the results as a benchmark to assist evaluation of the accuracy of ab initio calculations. Complexes containing [CeOH](2+) ligated by three- and four-acetone molecules were generated by electrospray ionization and characterized using wavelength-selective infrared multiple photon dissociation (IRMPD). The C[double bond, length as m-dash]O stretching frequency for the [CeOH(acetone)(4)](2+) species appeared at 1650 cm(-1) and was red-shifted by 90 cm(-1) compared to unligated acetone. The magnitude of this shift for the carbonyl frequency was even greater for the [CeOH(acetone)(3)](2+) complex: the IRMPD peak consisted of two dissociation channels, an initial elimination of acetone at 1635 cm(-1), and elimination of acetone concurrent with a charge separation producing [CeO(acetone)](+) at 1599 cm(-1), with the overall frequency centered at 1616 cm(-1). The increasing red shift observed as the number of acetone ligands decreases from four to three is consistent with transfer of more electron density per ligand in the less coordinated complexes. The lower frequency measured for the elimination/charge separation process is likely due to a combination of: (a) anharmonicity resulting from population of higher vibrational states, and (b) absorption by the initially formed photofragment [CeOH(acetone)(2)](2+). The C-C stretching frequency in the complexes is also influenced by coordination to the metal: it is blue-shifted compared to bare acetone, indicating a slight strengthening of the C-C bond in the complex, with the intensity of the absorption decreasing with decreasing ligation. Density functional theory (DFT) calculations using three different functionals (VWN, B3LYP, and PBE0) were used to predict the infrared spectra of the complexes. Calculated frequencies for the carbonyl stretch are within 40 cm(-1) of the IRMPD of the three-acetone complex measured using the single acetone loss, and within 60 cm(-1) of the measurement for the four-acetone complexes. The B3LYP functionals provided the best agreement with the measured spectra, with the VWN modestly lower and PBE0 modestly higher. The C-C stretching frequencies calculated using B3LYP are higher in energy than the measured values by approximately 30 cm(-1), and reproduce the observed trend which shows that the C-C stretching frequency decreases with increasing ligation. Agreement between C-C frequency and calculation was not as good using the VWN functional, but still within 70 cm(-1). The results provide an evaluation of changes in the acceptor properties of the metal center as ligands are added, and of the utility of DFT for modeling f-block coordination complexes.     

Making Mass Spectrometry See the Light: The Promises and Challenges of Cryogenic Infrared Ion Spectroscopy as a Bioanalytical Technique

The detailed chemical information contained in the vibrational spectrum of a cryogenically cooled analyte ion would, in principle, make infrared (IR) ion spectroscopy a gold standard technique for molecular identification in mass spectrometry. Despite this immense potential, there are considerable challenges in both instrumentation and methodology to overcome before the technique is analytically useful. Here, we discuss the promise of IR ion spectroscopy for small molecule analysis in the context of metabolite identification. Experimental strategies to address sensitivity constraints, poor overall duty cycle, and speed of the experiment are intimately tied to the development of a mass-selective cryogenic trap. Therefore, the most likely avenues for success, in the authors’ opinion, are presented here, alongside alternative approaches and some thoughts on data interpretation.

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Defying entropy: forming large head-to-tail macrocycles in the gas phase

Spectral fingerprints: Collision-induced dissociation (CID) of protonated peptides in the gas phase results in linear fragment ions with a five-membered oxazolone ring on their C-terminal side. Infrared spectroscopy confirms that smaller fragments adopt oxazolone structures. Conversely, in mid-sized and larger fragments an isomerization to "head-to-tail" macrocycles is observed (see picture).     

IRMPD spectroscopy of metal-ion/tryptophan complexes

Infrared multiple-photon dissociation (IRMPD) spectroscopy is employed to obtain detailed binding information on singly charged silver and alkali metal-ion/tryptophan complexes in the gas phase. For these complexes the presence of the salt bridge (i.e. zwitterionic) tautomer can be virtually excluded, except for perhaps a few percent in the case of Li+. Two low-energy structures having the charge solvation form are shown to be likely, where the metal cation is either in a tridentate N/O/Ring conformation or in a bidentate O/Ring conformation. These two structures can be conveniently discriminated and their relative quantities can be approximated by IR spectroscopy, based principally on diagnostic modes near approximately 1150 (N/O/Ring) and 1400 (O/Ring) cm(-1). Interestingly, the smaller cation complexes (i.e. Ag+ and Li+) display exclusively the N/O/Ring conformation, whereas the O/Ring conformer becomes progressively more abundant with increasing alkali metal size, eventually representing the majority of the ion population for Rb+ and Cs+. These spectroscopic findings are in excellent agreement with thermochemical density functional theory (DFT) calculations, giving support to the utility of high-level quantum-chemical calculations for such systems. Moreover, in contrast to other mass spectrometry-based techniques, IRMPD spectroscopy allows clear differentiation and semi-quantitative approximation of these metal-ligand binding motifs, thereby underlining its importance in thermochemical model benchmarking.