Electron capture dissociation and infrared multiphoton dissociation MS/MS of an N-glycosylated tryptic peptic to yield complementary sequence information
(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
- Håkansson, K ; Cooper, Helen J ; Emmett, Mark R ; Costello, Catherine E ; Marshall, A G and Nilsson, Carol LU
- publishing date
- 2001-09-15
- 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 (ACS)
- external identifiers
-
- scopus:0035884155
- pmid:11575803
- 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
- 2024-02-29 15:02:28
@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}},
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.}},
language = {{eng}},
month = {{09}},
number = {{18}},
pages = {{6--4530}},
publisher = {{The American Chemical Society (ACS)}},
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}},
doi = {{10.1021/ac0103470}},
volume = {{73}},
year = {{2001}},
}