Electron capture versus energetic dissociation of protein ions

Kruger, Nathan A.; Zubarev, Roman A.; Carpenter, Barry K.; Kelleher, Neil L.; Horn, David M.; McLafferty, Fred W.

doi:10.1016/s1387-3806(98)14260-4

Cited by 96 publications

(130 citation statements)

References 22 publications

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“…The digest was diluted to a final concentration of ϳ2 pmol/ L in methanol:water (50:50) and 1% formic acid. 15 N-isotopicnitrotyrosinelabelingofGPLEYGFAKGPLAK was completed by the electrochemical procedure described above, with the exception that Ͼ98% 15 N labeled sodium nitrate (Sigma-Aldrich, Poole, Dorset, UK) was used.…”

Section: Preparation Of Synthetic Peptidesmentioning

confidence: 99%

“…Apparently the ammonia loss is intrinsically linked with the nitrotyrosine radical anion formed following electron capture although the mechanism is unclear. 15 N-isotopic labeling of the nitro-group reveals that it is not the origin of the ammonia nitrogen (see Supplemental Figure 1, which can be found in the electronic version of this article). Presumably, the am- monia is lost from a protonated lysine side-chain which is interacting with the nitro-group.…”

Section: Loss Of Neutrals Observed Following Ecd Of Nitrated Peptidesmentioning

confidence: 99%

“…ECD differs from "slow-heating" MS/MS techniques [12], such as collisioninduced dissociation (CID) or infrared multi-photon dissociation (IRMPD) [13,14]. CID and IRMPD are thermal processes in which the lowest energy bonds are cleaved first [15]. The differences lead to different sets of fragmentation products being formed; ECD peptide backbone cleavage occurs at the N-C␣ bond, leading to c and z• (or c• and z) fragments [16,17], whereas CID and IRMPD peptide backbone cleavage occurs at N-C O bonds producing b and y ions [18].…”

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confidence: 99%

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Electron capture dissociation mass spectrometry of tyrosine nitrated peptides

Jones

Mikhailov

Iniesta

et al. 2010

J. Am. Soc. Mass Spectrom.

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In vivo protein nitration is associated with many disease conditions that involve oxidative stress and inflammatory response. The modification involves addition of a nitro group at the position ortho to the phenol group of tyrosine to give 3-nitrotyrosine. To understand the mechanisms and consequences of protein nitration, it is necessary to develop methods for identification of nitrotyrosine-containing proteins and localization of the sites of modification. Here, we have investigated the electron capture dissociation (ECD) and collision-induced dissociation (CID) behavior of 3-nitrotyrosine-containing peptides. The presence of nitration did not affect the CID behavior of the peptides. For the doubly-charged peptides, addition of nitration severely inhibited the production of ECD sequence fragments. However, ECD of the triply-charged nitrated peptides resulted in some singly-charged sequence fragments. ECD of the nitrated peptides is characterized by multiple losses of small neutral species including hydroxyl radicals, water and ammonia. The origin of the neutral losses has been investigated by use of activated ion (AI) ECD. Loss of ammonia appears to be the result of non-covalent interactions between the nitro group and protonated lysine side-chains. (J Am Soc Mass Spectrom 2010, 21, 268 -277)

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Section: Preparation Of Synthetic Peptidesmentioning

confidence: 99%

Section: Loss Of Neutrals Observed Following Ecd Of Nitrated Peptidesmentioning

confidence: 99%

mentioning

confidence: 99%

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Electron capture dissociation mass spectrometry of tyrosine nitrated peptides

Jones

Mikhailov

Iniesta

et al. 2010

J. Am. Soc. Mass Spectrom.

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“…A far larger proportion of interresidue bonds is cleaved using the related techniques of ECD [23][24][25][26][27]38,[59][60][61][62] and ETD [19,20,28,29]. In ECD, a low energy (≪1 eV) electron is captured by a multiply protonated protein, (M + nH) n+ , to form a reduced (M + nH) (n−1)+• radical ion; for ETD the electron is supplied by an aromatic radical anion.…”

Section: Methods For Dissociation Of Multiply-charged Biomolecular Ionsmentioning

confidence: 99%

“…This ~5 eV energy gain also makes the <1 eV differences in interresidue bond dissociation energies have far less influence on product abundances. Thus ECD and ETD backbone cleavage is much less affected by the identity of the amino acids (except for proline, vide infra) framing the cleavage site, and in this way it provides more extensive sequence coverage and PTM location specificity [25,59].…”

Section: Methods For Dissociation Of Multiply-charged Biomolecular Ionsmentioning

confidence: 99%

Top-down identification and characterization of biomolecules by mass spectrometry

Breuker

Jin

Han

et al. 2008

J. Am. Soc. Mass Spectrom.

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112

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The most widely used modern mass spectrometers face severe performance limitations with molecules larger than a few kDa. For far larger biomolecules, a common practice has been to break these up chemically or enzymatically into fragments that are sufficiently small for the instrumentation available. With its many sophisticated recent enhancements, this "bottom-up" approach has proved highly valuable, such as for the rapid, routine identification and quantitation of DNA-predicted proteins in complex mixtures. Characterization of smaller molecules, however, has always measured the mass of the molecule and then that of its fragments. This "top-down" approach has been made possible for direct analysis of large biomolecules by the uniquely high (>10 5 ) mass resolving power and accuracy (~1 ppm) of the Fourier-transform mass spectrometer. For complex mixtures, isolation of a single component's molecular ions for MS/MS not only gives biomolecule identifications of far higher reliability, but directly characterizes sequence errors and posttranslational modifications. Protein sizes amenable for current MS/MS instrumentation are increased by a "middle-down" approach in which limited proteolysis forms large (e.g., 10 kDa) polypeptides that are then subjected to the top-down approach, or by "prefolding dissociation". The latter, which extends characterization to proteins >200 kDa, was made possible by greater understanding of how molecular ion tertiary structure evolves in the gas phase.

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Ion trap collisional activation of disulfide linkage intact and reduced multiply protonated polypeptides†

Stephenson

Cargile

McLuckey

1999

Rapid Commun. Mass Spectrom.

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The presence of disulfide linkages in multiply charged polypeptide ions tends to inhibit the formation of structurally informative product ions under conventional quadrupole ion trap collisional activation conditions. In particular, fragmentation that requires two cleavages (i.e., cleavage of a disulfide linkage and a peptide linkage) is strongly suppressed. Reduction of the disulfide linkage(s) by use of dithiothreitol yields parent ions upon electrospray without this complication. Far richer structural information is revealed by ion trap collisional activation of the disulfide-reduced species than from the native species. These observations are illustrated with doubly protonated native and reduced somatosin, the [M + 5H](5+) ion of native bovine insulin and the [M + 4H](4+) and [M + 3H](3+) ions of the B-chain of bovine insulin produced by reduction of the disulfide linkages in insulin, and the [M + 11H](11+) ion of native chicken lysozyme and the [M + 11H](11+) and [M + 14H](14+) ions of reduced lysozyme. In each case, the product ions produced by ion trap collisional activation were subjected to ion/ion proton transfer reactions to facilitate interpretation of the product ion spectra. These studies clearly suggest that the identification of polypeptides with one or more disulfide linkages via application of ion trap collisional activation to the multiply charged parent ions formed directly by electrospray could be problematic. Means for cleaving the disulfide linkage, such as reduction by dithiothreitol prior to electrospray, are therefore desirable in these cases.

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Electron capture versus energetic dissociation of protein ions

Cited by 96 publications

References 22 publications

Electron capture dissociation mass spectrometry of tyrosine nitrated peptides

Electron capture dissociation mass spectrometry of tyrosine nitrated peptides

Top-down identification and characterization of biomolecules by mass spectrometry

Ion trap collisional activation of disulfide linkage intact and reduced multiply protonated polypeptides†

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