Charge state dependent fragmentation of gaseous protein ions in a quadrupole ion trap: bovine ferri-, ferro-, and apo-cytochrome c

Engel, Brian J.; Pan, Peng; Reid, Gavin E.; Wells, Justin; McLuckey, Scott A.

doi:10.1016/s1387-3806(02)00562-6

Cited by 54 publications

(68 citation statements)

References 65 publications

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“…Future linear trap instruments designed for this purpose will undoubtedly contain additional segments plus controls for superposition of multiple RF, ac, and dc fields. These features will allow implementation of even more extensive ion͞ion manipulations, which almost certainly will include fully independent isolation of precursor and reagent ion clusters plus the experiments pioneered by McLuckey et al (40)(41)(42)(43)(44)(45)(46) for charge-state reduction and gas-phase (charge-state) concentration on the QIT instrument. Additionally, we envision implementing techniques that prevent ETD products from undergoing subsequent ion͞ion reactions that lead to either neutralization of charge or formation of additional fragments.…”

Section: Instrumentationmentioning

confidence: 99%

Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry

Syka

Coon

Schroeder

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

2,230

2,264

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Peptide sequence analysis using a combination of gas-phase ion͞ion chemistry and tandem mass spectrometry (MS͞MS) is demonstrated. Singly charged anthracene anions transfer an electron to multiply protonated peptides in a radio frequency quadrupole linear ion trap (QLT) and induce fragmentation of the peptide backbone along pathways that are analogous to those observed in electron capture dissociation. Modifications to the QLT that enable this ion͞ion chemistry are presented, and automated acquisition of high-quality, single-scan electron transfer dissociation MS͞MS spectra of phosphopeptides separated by nanoflow HPLC is described.electron capture dissociation ͉ fragmentation ͉ ion͞ion reactions ͉ charge transfer ͉ ion trap S ix years ago, McLafferty and coworkers (1) introduced a unique method for peptide͞protein ion fragmentation: electron capture dissociation (ECD). In this method, multiply protonated peptides or proteins are confined in the Penning trap of a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer and exposed to electrons with near-thermal energies. Capture of a thermal electron by a protonated peptide is exothermic by Ϸ6 eV (1 eV ϭ 1.602 ϫ 10 Ϫ19 J) and causes the peptide backbone to fragment by a nonergodic process, e.g., one that does not involve intramolecular vibrational energy redistribution (2-5). One pathway for this process involves generation of an odd-electron hypervalent species (RNH 3 • ) that dissociates to produce RNH 2 and a hydrogen radical (6). As shown in Fig to an amide nitrogen, a secondary pathway, leads to the formation of carbon monoxide plus a homologous series of complementary fragment ions of types a and y. Subtraction of the m͞z values for the fragments within a given ion series that differ by a single amino acid affords the mass and thus the identity of the extra residue in the larger of the two fragments. The complete amino acid sequence of a peptide is deduced by extending this process to all homologous pairs of fragments within a particular ion series.Because ECD occurs along the peptide backbone in a sequence-independent manner, preserves posttranslational modifications (PTMs) (7-14), and can be implemented on a millisecond time scale with precursor-to-product ion conversion efficiencies that approach 30% (15-21), it has become the technique of choice for the analysis of peptide and proteins with FTICR mass spectrometers (22-28). Unfortunately, ECD in its most efficient form requires that the precursor sample ions be immersed in a dense population of near-thermal electrons. Emulating these conditions in the instruments used most commonly for peptide and protein analyses, those that trap ions with radio frequency (RF) electrostatic fields rather than with static magnetic and electric fields, remains technically challenging. Thermal electrons introduced into the RF fields of RF 3D quadrupole ion trap (QIT), quadrupole time-of-flight, or RF linear 2D quadrupole ion trap (QLT) instruments maintain their thermal energy only for a fraction of a micros...

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Section: Instrumentationmentioning

confidence: 99%

Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry

Syka

Coon

Schroeder

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

2,230

2,264

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“…Ion/ion reactions provide efficient means for manipulating the identities of gas-phase ions after their initial formation [3,4], including altering ion polarity and absolute charge. Applications of charge manipulation via ion/ ion reactions include: mixture analysis [5][6][7][8], especially with the application of the "ion parking" technique for gas-phase concentration and charge state purification [9], the formation of ions that cannot be directly produced by ESI for subsequent tandem mass spectrometry studies [10], and the reduction of the product ion charge states to singly and doubly charged so as to simplify the interpretation of product ion spectra [11][12][13][14][15][16]. Electron-transfer ion/ion reactions have recently been described as a means of inducing structurally informative dissociation, giving rise to cleavages analogous to those noted in electron capture dissociation [17][18][19][20][21].…”

mentioning

confidence: 99%

Pulsed dual electrospray ionization for In/In reactions

Xia

Liang

McLuckey

2005

J. Am. Soc. Mass Spectrom.

116

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A pulsed dual electrospray ionization source has been developed to generate positive and negative ions for subsequent ion/ion reaction experiments. The two sprayers, typically a nano-electrospray emitter for analytes and an electrospray emitter for reagents, are positioned in a parallel fashion close to the sampling orifice of a triple quadrupole/linear ion trap tandem mass spectrometer (Sciex Q TRAP). The potentials applied to each sprayer are alternately pulsed so that ions of opposite polarity are generated separately in time. Ion/ion reactions take place after ions of each polarity are sequentially injected into a high-pressure linear ion trap, where axial trapping is effected by applying an auxiliary radio frequency voltage to the end lenses. The pulsed dual electrospray source allows optimization of each sprayer and can be readily coupled to any spray interface with no need for instrument modifications, provided the potentials required to transmit the ion polarity of interest can be alternated in synchrony with the emitter potentials. Ion/ion reaction examples such as charge reduction of multiply charged protein ions, charge inversion of peptides ions, and protein-protein complex formation are given to illustrate capabilities of the pulsed dual electrospray source in the study of gas-phase ion/ion chemistry. T he advent of electrospray ionization (ESI) mass spectrometry has revolutionized the analysis of many biologically important macromolecules [1,2]. Both the polarity and the magnitude of the charge associated with an ionized biomolecule play important roles in the use of tandem mass spectrometry for the identification and structural characterization of the biomolecule from which the ions were derived. Ion/ion reactions provide efficient means for manipulating the identities of gas-phase ions after their initial formation [3,4], including altering ion polarity and absolute charge. Applications of charge manipulation via ion/ ion reactions include: mixture analysis [5][6][7][8], especially with the application of the "ion parking" technique for gas-phase concentration and charge state purification [9], the formation of ions that cannot be directly produced by ESI for subsequent tandem mass spectrometry studies [10], and the reduction of the product ion charge states to singly and doubly charged so as to simplify the interpretation of product ion spectra [11][12][13][14][15][16]. Electron-transfer ion/ion reactions have recently been described as a means of inducing structurally informative dissociation, giving rise to cleavages analogous to those noted in electron capture dissociation [17][18][19][20][21]. In this regard, ion/ion reactions play a role as a probe of primary structure.Electrodynamic ion traps, either a quadrupole ion trap or a linear ion trap (LIT), are particularly useful reaction vessels for ion/ion reactions because of their ability to store oppositely-charged ions simultaneously [22] as well as their ion isolation and MS n functionalities [23]. Most ion/ion reaction studies involving m...

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“…Other groups have recently shown that the information obtained from CID data obtained from different charge states can vary substantially and that it is useful to examine a number of charge states in order to extract detailed information about amino acid sequence across large portions of the protein [21,22]. In favorable cases a single IMS-CID/MS frame may contain fragmentation for many charge states; thus, the approach is suitable for high-throughput characterization.…”

Section: Demonstration Of High-field Lc/ims-cid/ms For a Simple Protementioning

confidence: 99%

“…For example, electrospray ionization (ESI) usually produces multiple charge states. Because fragment patterns from different charge states are often complementary for obtaining high-levels of sequence coverage and the positions of modifications, it is desirable to mass-select and fragment many charge states [21,22]. However, in practice it is often not possible to select, activate, and detect fragments from multiple charge states when experiments are carried out on-line because components may be present for analysis for only fractions of a minute.…”

mentioning

confidence: 99%

Nanoflow LC/IMS-MS and LC/IMS-CID/MS of protein mixtures

Sowell

Koeniger

Valentine

et al. 2004

J. Am. Soc. Mass Spectrom.

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A simple ion trap/ion mobility/time-of-flight (TOF) mass spectrometer has been coupled with nanoflow liquid chromatography to examine the feasibility of analyzing mixtures of intact proteins. In this approach proteins are separated using reversed-phase chromatography. As components elute from the column, they are electrosprayed into the gas phase and separated again in a drift tube prior to being dispersed and analyzed in a TOF mass spectrometer. The mobilities of ions through a buffer gas depend upon their collision cross sections and charge states; separation based on these gas-phase parameters provides a new means of simplifying mass spectra and characterizing mixtures. Additionally it is possible to induce dissociation at the exit of the drift tube and examine the fragmentation patterns of specific protein ion charge states and conformations. The approach is demonstrated by examining a simple threecomponent mixture containing ubiquitin, cytochrome c, and myoglobin and several larger prepared protein mixtures. The potential of this approach for use in proteomic applications is considered. (J Am Soc Mass Spectrom 2004, 15, 1341-1353

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Charge state dependent fragmentation of gaseous protein ions in a quadrupole ion trap: bovine ferri-, ferro-, and apo-cytochrome c

Cited by 54 publications

References 65 publications

Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry

Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry

Pulsed dual electrospray ionization for In/In reactions

Nanoflow LC/IMS-MS and LC/IMS-CID/MS of protein mixtures

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