Characterization and optimization of electron detachment dissociation Fourier transform ion cyclotron resonance mass spectrometry

Yang, Jinggang; Håkansson, Kristina

doi:10.1016/j.ijms.2008.05.036

Cited by 22 publications

(21 citation statements)

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“…An alternative method that was reported to actually give higher yields of sequenceinformative fragment ions from more highly and negatively charged precursors [28,29] is electron detachment dissociation (EDD), which was introduced by Zubarev and coworkers as a new technique for dissociation of peptide anions [30]. In EDD, bombardment of multiply deprotonated peptides [31][32][33], oligonucleotides [29, 34 -38], or oligosaccharides [28, 39 -43] with Ͼ10 eV electrons results in electron detachment, producing (M Ϫ nH) (n-1)-• radical ions that can undergo backbone dissociation [37,38].…”

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confidence: 99%

Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

Taucher

Breuker

2010

J. Am. Soc. Mass Spectrom.

109

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We have studied the effect of solution additives on hydrolysis and charge state distribution in ESI MS of RNA. Lower and higher charge state ions can be electrosprayed from solutions containing 25 mM piperidine/25 mM imidazole and 1% vol. triethylamine, respectively, with base-catalyzed hydrolysis rates that are sufficiently slow to perform MS/MS experiments. These lower and higher charge state ions are suitable as precursors for CAD and EDD, respectively. We demonstrate nearly complete sequence coverage for 61 nt RNA dissociated by CAD, and 34 nt RNA dissociated by EDD, and suggest a mechanism for backbone fragmentation in EDD of RNA. (J Am Soc Mass Spectrom 2010, 21, 918 -929) © 2010 American Society for Mass Spectrometry T he "top-down" approach is increasingly being applied in mass spectrometry (MS) studies of proteins [1][2][3][4][5], using mostly collisionally activated dissociation (CAD) [6 -8], electron capture dissociation (ECD) [9 -12], or electron-transfer dissociation (ETD) [13] for generation of sequence-informative fragment ions from backbone cleavage in multiply protonated precursor ions. First examples of top-down mass spectrometry of smaller (up to 25 nt) multiply deprotonated desoxyribonucleic acids (DNA) were reported in the early 1990s [14 -16], and the concept was soon extended to larger (up to 108 nt) DNA [17].For ribonucleic acids (RNAs), the development and application of mass spectrometry-based methodology has been much slower than for DNA [18], possibly because the significance of RNA in regulation of gene expression was recognized only recently [19]. Topdown MS has been applied to study RNA modified by structural probes [20], for characterization of conserved domains of the HIV-1 packaging signal RNA [21], for investigating aptamer/ligand complexes [22], and binding of antibiotics to ribosomal RNA subdomains [23]. However, sequencing by top-down mass spectrometry of RNA Ͼ 20 nt has been demonstrated only recently, as discussed below.Collisionally activated dissociation of multiply deprotonated RNA from electrospray ionization (ESI) [24] has lately provided full sequence coverage for small interfering RNA (siRNA) [25] and a riboswitch aptamer domain sequence [26] consisting of 21 and 34 nucleotides (nt), respectively. Mass spectral quality, particularly with regard to undesired secondary fragmentation, was shown to critically depend on precursor ion charge [25,27] and collisional cooling of primary fragments [26]. Our rationale for the decrease in secondary fragmentation was that reduced net charge and collisional cooling minimize the internal energy of fragment ions from primary backbone cleavage, which makes them less prone to secondary fragmentation [26]. In our study of 34 nt RNA, we used ions of relatively low net charge, (M Ϫ 7H) 7Ϫ , which were electrosprayed from acidified solutions [26].However, mass spectrometer performance generally decreases with increasing mass-to-charge ratio, i.e., lower ion charge. An alternative method that was reported to actually give higher yields...

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confidence: 99%

Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

Taucher

Breuker

2010

J. Am. Soc. Mass Spectrom.

109

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“…For EDD, the cathode heating current was kept at 2.0 A and the cathode voltage was pulsed to a bias voltage of − 30 to − 35 V [34] for 1 s. An extraction lens located in between the cathode and the ICR cell was kept − 0.8 to − 1.0 V higher than the cathode bias voltage. An electron current of a few μA is optimum for EDD [75]. IRMPD was performed with a laser power of 10 W with firing times ranging from 0.25–1s.…”

Section: Methodsmentioning

confidence: 99%

Electron detachment dissociation of fluorescently labeled sialylated oligosaccharides

Zhou

Håkansson

2011

Electrophoresis

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We explored the application of electron detachment dissociation (EDD) and infrared multiphoton dissociation (IRMPD) tandem mass spectrometry to fluorescently labeled sialylated oligosaccharides. Standard sialylated oligosaccharides and a sialylated N-linked glycan released from human transferrin were investigated. EDD yielded extensive glycosidic cleavages and cross-ring cleavages in all cases studied, consistently providing complementary structural information compared to IRMPD. Neutral losses and satellite ions such as C – 2H ions were also observed following EDD. In addition, we examined the influence of different fluorescent labels. The acidic label 2-aminobenzoic acid (2-AA) enhanced signal abundance in negative-ion mode. However, few cross-ring fragments were observed for 2-AA labeled oligosaccharides. The neutral label 2-aminobenzamide (2-AB) resulted in more cross-ring cleavages compared to 2-AA labeled species, but not as extensive fragmentation as for native oligosaccharides, likely resulting from altered negative charge locations from introduction of the fluorescent tag.

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“…All experiments were performed with a 7-T FT-ICR mass spectrometer with a quadrupole front end (APEX-Q, Bruker Daltonics, Billerica, MA, USA) [44]. Oligosaccharide solutions were infused via an external Apollo II electrospray ion source at a flow rate of 60 μL/h with the assistance of N 2 nebulizing gas.…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…Precursor ions were further isolated in the ICR cell using correlated harmonic excitation fields (CHEF) [46]. An indirectly heated hollow dispenser cathode [47] was used to perform EDD with a constant heating current of 2.0 A and a pulsed cathode bias voltage that varied between -20 and -35 V for 1-2 s. A 6 mm diameter lens electrode located immediately in front of the cathode was held 0.4-1.0 V higher than the bias voltage to regulate the number of electrons entering the ICR cell [44]. External CAD was performed in a hexapole following mass selective ion accumulation with argon as collision gas.…”

Section: Mass Spectrometrymentioning

confidence: 99%

Electron Detachment Dissociation of Underivatized Chloride-Adducted Oligosaccharides

Kornacki

Adamson

Håkansson

2012

J. Am. Soc. Mass Spectrom.

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Chloride anion attachment has previously been shown to aid determination of saccharide anomeric configuration and generation of linkage information in negative ion post-source decay MALDI tandem mass spectrometry. Here, we employ electron detachment dissociation (EDD) and collision activated dissociation (CAD) for the structural characterization of underivatized oligosaccharides bearing a chloride ion adduct. Both neutral and sialylated oligosaccharides are examined, including maltoheptaose, an asialo biantennary glycan (NA2), disialylacto-N-tetraose (DSLNT), and two LS tetrasaccharides (LSTa and LSTb). Gas-phase chloride-adducted species are generated by negative ion mode electrospray ionization. EDD and CAD spectra of chlorideadducted oligosaccharides are compared to the corresponding spectra for doubly deprotonated species not containing a chloride anion to assess the role of chloride adduction in the stimulation of alternative fragmentation pathways and altered charge locations allowing detection of additional product ions. In all cases, EDD of singly chloridated and singly deprotonated species resulted in an increase in observed cross-ring cleavages, which are essential to providing saccharide linkage information. Glycosidic cleavages also increased in EDD of chlorideadducted oligosaccharides to reveal complementary structural information compared to traditional (non-chloride-assisted) EDD and CAD. Results indicate that chloride adduction is of interest in alternative anion activation methods such as EDD for oligosaccharide structural characterization.

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Characterization and optimization of electron detachment dissociation Fourier transform ion cyclotron resonance mass spectrometry

Cited by 22 publications

References 34 publications

Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

Electron detachment dissociation of fluorescently labeled sialylated oligosaccharides

Electron Detachment Dissociation of Underivatized Chloride-Adducted Oligosaccharides

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