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Electron capture dissociation and infrared multiphoton dissociation MS/MS of an N-glycosylated tryptic peptic to yield complementary sequence information

Håkansson, K; Cooper, Helen J; Emmett, Mark R; Costello, Catherine E; Marshall, A G and Nilsson, Carol LU (2001) In Analytical Chemistry 73(18). p.6-4530
Abstract

Glycoproteins are a functionally important class of biomolecules for which structural elucidation presents a challenge. Fragmentation of N-glycosylated peptides, employing collisionally activated dissociation, typically yields product ions that result from dissociation at glycosidic bonds, with little occurrence of dissociation at peptide backbone sites. We have applied two dissociation techniques, electron capture dissociation (ECD) and infrared multiphoton dissociation (IRMPD), in a 7-T Fourier transform ion cyclotron resonance mass spectrometer, in the investigation of an N-glycosylated peptide from an unfractionated tryptic digest of the lectin of the coral tree, Erythrina corallodendron. ECD provided c and z. ions derived from the... (More)

Glycoproteins are a functionally important class of biomolecules for which structural elucidation presents a challenge. Fragmentation of N-glycosylated peptides, employing collisionally activated dissociation, typically yields product ions that result from dissociation at glycosidic bonds, with little occurrence of dissociation at peptide backbone sites. We have applied two dissociation techniques, electron capture dissociation (ECD) and infrared multiphoton dissociation (IRMPD), in a 7-T Fourier transform ion cyclotron resonance mass spectrometer, in the investigation of an N-glycosylated peptide from an unfractionated tryptic digest of the lectin of the coral tree, Erythrina corallodendron. ECD provided c and z. ions derived from the peptide backbone, with no observed loss of sugars. Cleavage at 11 of 15 backbone amine bonds was observed. The lack of cleavage at sites located close to the glycosylated asparagine residue may result from steric blocking by the glycan. IRMPD provided abundant fragment ions, primarily through dissociation at glycosidic linkages. The monosaccharide composition and the presence of three glycan branch sites could be determined from the IRMPD fragments. The two types of spectra, obtained with the same instrument, thus provide complementary structural information about the glycopeptide. The current result extends the applicability of ECD for glycopeptide analysis to N-glycosylated peptides and to peptides containing branched, highly substituted glycans.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Electrons, Glycoproteins, Glycosylation, Lectins, Peptides, Sequence Analysis, Protein, Spectrometry, Mass, Electrospray Ionization, Spectroscopy, Fourier Transform Infrared, Trypsin, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.
in
Analytical Chemistry
volume
73
issue
18
pages
6 - 4530
publisher
The American Chemical Society
external identifiers
  • scopus:0035884155
ISSN
0003-2700
DOI
10.1021/ac0103470
language
English
LU publication?
no
id
d36335cc-8501-4fd9-b23b-5f7bd5be0f0f
date added to LUP
2017-05-16 10:42:52
date last changed
2018-07-29 04:26:28
@article{d36335cc-8501-4fd9-b23b-5f7bd5be0f0f,
  abstract     = {<p>Glycoproteins are a functionally important class of biomolecules for which structural elucidation presents a challenge. Fragmentation of N-glycosylated peptides, employing collisionally activated dissociation, typically yields product ions that result from dissociation at glycosidic bonds, with little occurrence of dissociation at peptide backbone sites. We have applied two dissociation techniques, electron capture dissociation (ECD) and infrared multiphoton dissociation (IRMPD), in a 7-T Fourier transform ion cyclotron resonance mass spectrometer, in the investigation of an N-glycosylated peptide from an unfractionated tryptic digest of the lectin of the coral tree, Erythrina corallodendron. ECD provided c and z. ions derived from the peptide backbone, with no observed loss of sugars. Cleavage at 11 of 15 backbone amine bonds was observed. The lack of cleavage at sites located close to the glycosylated asparagine residue may result from steric blocking by the glycan. IRMPD provided abundant fragment ions, primarily through dissociation at glycosidic linkages. The monosaccharide composition and the presence of three glycan branch sites could be determined from the IRMPD fragments. The two types of spectra, obtained with the same instrument, thus provide complementary structural information about the glycopeptide. The current result extends the applicability of ECD for glycopeptide analysis to N-glycosylated peptides and to peptides containing branched, highly substituted glycans.</p>},
  author       = {Håkansson, K and Cooper, Helen J and Emmett, Mark R and Costello, Catherine E and Marshall, A G and Nilsson, Carol},
  issn         = {0003-2700},
  keyword      = {Electrons,Glycoproteins,Glycosylation,Lectins,Peptides,Sequence Analysis, Protein,Spectrometry, Mass, Electrospray Ionization,Spectroscopy, Fourier Transform Infrared,Trypsin,Journal Article,Research Support, Non-U.S. Gov't,Research Support, U.S. Gov't, Non-P.H.S.,Research Support, U.S. Gov't, P.H.S.},
  language     = {eng},
  month        = {09},
  number       = {18},
  pages        = {6--4530},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Electron capture dissociation and infrared multiphoton dissociation MS/MS of an N-glycosylated tryptic peptic to yield complementary sequence information},
  url          = {http://dx.doi.org/10.1021/ac0103470},
  volume       = {73},
  year         = {2001},
}