Tags

Type your tag names separated by a space and hit enter

Electron transfer dissociation of N-glycopeptides: loss of the entire N-glycosylated asparagine side chain.
Rapid Commun Mass Spectrom. 2007; 21(6):1053-61.RC

Abstract

The recently introduced electron transfer dissociation (ETD) technique opens new possibilities for the structural characterization of glycoproteins at the glycopeptide level. In this report, we investigate the ETD mass spectra of tryptic N-glycopeptides of the model glycoprotein horseradish peroxidase (HRP). Multiply protonated N-glycopeptides obtained by electrospray ionization were subjected to ETD. Fragment ions obtained by ETD were further analyzed by collision-induced dissociation (CID) (MS(3)) for their unambiguous structural assignment. The following fragmentation features were revealed: (1) c- and z-type peptide backbone cleavages were observed with retention of the intact glycan moiety revealing peptide sequence, glycan attachment site, and glycan mass; (2) to a lesser extent, glycosidic bond cleavages were registered with retention of the intact peptide sequence; and (3) a range of amino acid side chain losses did occur. Remarkably, the loss of the complete N-glycosylated asparagine side chain was observed. This loss of the glycan-modified side chain helps with the structural characterization of glycopeptides by allowing the facile deduction and verification of the glycan mass and the nature of the amino acid residue at the glycan attachment site. Importantly, informative ETD spectra were obtained in this study by reversed-phase nano-liquid chromatography (LC) coupled online to a radio-frequency (rf) quadrupole ion trap (QIT) mass spectrometer with alternating acquisition of CID and ETD mass spectra from an automatically selected set of precursors (data-dependent mode). Thus, our study brings nano-LC/QIT-MS(n) with CID and ETD to the fore as a powerful technique for glycoproteomics at the glycopeptide level.

Authors+Show Affiliations

Biomolecular Mass Spectrometry Unit, Department of Parasitology, Center of Infectious Diseases, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17311219

Citation

Catalina, M Isabel, et al. "Electron Transfer Dissociation of N-glycopeptides: Loss of the Entire N-glycosylated Asparagine Side Chain." Rapid Communications in Mass Spectrometry : RCM, vol. 21, no. 6, 2007, pp. 1053-61.
Catalina MI, Koeleman CA, Deelder AM, et al. Electron transfer dissociation of N-glycopeptides: loss of the entire N-glycosylated asparagine side chain. Rapid Commun Mass Spectrom. 2007;21(6):1053-61.
Catalina, M. I., Koeleman, C. A., Deelder, A. M., & Wuhrer, M. (2007). Electron transfer dissociation of N-glycopeptides: loss of the entire N-glycosylated asparagine side chain. Rapid Communications in Mass Spectrometry : RCM, 21(6), 1053-61.
Catalina MI, et al. Electron Transfer Dissociation of N-glycopeptides: Loss of the Entire N-glycosylated Asparagine Side Chain. Rapid Commun Mass Spectrom. 2007;21(6):1053-61. PubMed PMID: 17311219.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Electron transfer dissociation of N-glycopeptides: loss of the entire N-glycosylated asparagine side chain. AU - Catalina,M Isabel, AU - Koeleman,Carolien A M, AU - Deelder,André M, AU - Wuhrer,Manfred, PY - 2007/2/22/pubmed PY - 2007/5/10/medline PY - 2007/2/22/entrez SP - 1053 EP - 61 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 21 IS - 6 N2 - The recently introduced electron transfer dissociation (ETD) technique opens new possibilities for the structural characterization of glycoproteins at the glycopeptide level. In this report, we investigate the ETD mass spectra of tryptic N-glycopeptides of the model glycoprotein horseradish peroxidase (HRP). Multiply protonated N-glycopeptides obtained by electrospray ionization were subjected to ETD. Fragment ions obtained by ETD were further analyzed by collision-induced dissociation (CID) (MS(3)) for their unambiguous structural assignment. The following fragmentation features were revealed: (1) c- and z-type peptide backbone cleavages were observed with retention of the intact glycan moiety revealing peptide sequence, glycan attachment site, and glycan mass; (2) to a lesser extent, glycosidic bond cleavages were registered with retention of the intact peptide sequence; and (3) a range of amino acid side chain losses did occur. Remarkably, the loss of the complete N-glycosylated asparagine side chain was observed. This loss of the glycan-modified side chain helps with the structural characterization of glycopeptides by allowing the facile deduction and verification of the glycan mass and the nature of the amino acid residue at the glycan attachment site. Importantly, informative ETD spectra were obtained in this study by reversed-phase nano-liquid chromatography (LC) coupled online to a radio-frequency (rf) quadrupole ion trap (QIT) mass spectrometer with alternating acquisition of CID and ETD mass spectra from an automatically selected set of precursors (data-dependent mode). Thus, our study brings nano-LC/QIT-MS(n) with CID and ETD to the fore as a powerful technique for glycoproteomics at the glycopeptide level. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/17311219/Electron_transfer_dissociation_of_N_glycopeptides:_loss_of_the_entire_N_glycosylated_asparagine_side_chain_ L2 - https://doi.org/10.1002/rcm.2929 DB - PRIME DP - Unbound Medicine ER -