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Structural analysis of O-glycopeptides employing negative- and positive-ion multi-stage mass spectra obtained by collision-induced and electron-capture dissociations in linear ion trap time-of-flight mass spectrometry.
Rapid Commun Mass Spectrom. 2007; 21(5):691-8.RC

Abstract

Structural analyses of various glycans attached to proteins and peptides are highly desirable for elucidating their biological roles. An approach based on mass spectrometry (MS) combining both collision-induced dissociation (CID) and electron-capture dissociation (ECD) in the positive- and negative-ion modes has been proposed as a simple and direct method of assigning an O-glycan without releasing it from the peptide and of determining the amino acid sequence of the peptide and glycosylation site. The instrument used is an electrospray ionization (ESI) linear ion trap (LIT) time-of-flight (TOF) mass spectrometer with tandem LITs for CID by He gas and ECD. The proposed approach was tested with two synthetic O-glycopeptides binding a sialyl Lewis x (sLe(x)) oligosaccharide and a 3'-sialyl N-acetyllactosamine (3'-SLN) on a serine (S) residue. In the negative-ion mode, the CID MS(2) spectra of O-glycopeptides showed a relatively abundant glycoside-bond cleavage between the core N-acetylglucosamine (GlcNAc) and serine (S) that yields deprotonated C(3)-type fragment ions of O-glycan and deprotonated Z(0)-type peptide ions. The structure of the sLe(x) (3'-SLN) oligosaccharide was simply assigned by comparing the CID MS(3) spectrum derived from the C(3)-type fragment ion with the CID MS(2) spectra of the sLe(x) and sLe(a) (3'- and 6'-SLN) standards (i.e., negative-ion MS(n) spectral matching). The amino acid sequence of the peptide including the glycosylation site was determined from the ECD MS(2) spectrum in the positive-ion mode.

Authors+Show Affiliations

Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan. deguchi@glyco.sci.hokudai.ac.jpNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17279605

Citation

Deguchi, Kisaburo, et al. "Structural Analysis of O-glycopeptides Employing Negative- and Positive-ion Multi-stage Mass Spectra Obtained By Collision-induced and Electron-capture Dissociations in Linear Ion Trap Time-of-flight Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 21, no. 5, 2007, pp. 691-8.
Deguchi K, Ito H, Baba T, et al. Structural analysis of O-glycopeptides employing negative- and positive-ion multi-stage mass spectra obtained by collision-induced and electron-capture dissociations in linear ion trap time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 2007;21(5):691-8.
Deguchi, K., Ito, H., Baba, T., Hirabayashi, A., Nakagawa, H., Fumoto, M., Hinou, H., & Nishimura, S. (2007). Structural analysis of O-glycopeptides employing negative- and positive-ion multi-stage mass spectra obtained by collision-induced and electron-capture dissociations in linear ion trap time-of-flight mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 21(5), 691-8.
Deguchi K, et al. Structural Analysis of O-glycopeptides Employing Negative- and Positive-ion Multi-stage Mass Spectra Obtained By Collision-induced and Electron-capture Dissociations in Linear Ion Trap Time-of-flight Mass Spectrometry. Rapid Commun Mass Spectrom. 2007;21(5):691-8. PubMed PMID: 17279605.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structural analysis of O-glycopeptides employing negative- and positive-ion multi-stage mass spectra obtained by collision-induced and electron-capture dissociations in linear ion trap time-of-flight mass spectrometry. AU - Deguchi,Kisaburo, AU - Ito,Hiroki, AU - Baba,Takashi, AU - Hirabayashi,Atsumu, AU - Nakagawa,Hiroaki, AU - Fumoto,Masataka, AU - Hinou,Hiroshi, AU - Nishimura,Shin-Ichiro, PY - 2007/2/7/pubmed PY - 2007/5/2/medline PY - 2007/2/7/entrez SP - 691 EP - 8 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 21 IS - 5 N2 - Structural analyses of various glycans attached to proteins and peptides are highly desirable for elucidating their biological roles. An approach based on mass spectrometry (MS) combining both collision-induced dissociation (CID) and electron-capture dissociation (ECD) in the positive- and negative-ion modes has been proposed as a simple and direct method of assigning an O-glycan without releasing it from the peptide and of determining the amino acid sequence of the peptide and glycosylation site. The instrument used is an electrospray ionization (ESI) linear ion trap (LIT) time-of-flight (TOF) mass spectrometer with tandem LITs for CID by He gas and ECD. The proposed approach was tested with two synthetic O-glycopeptides binding a sialyl Lewis x (sLe(x)) oligosaccharide and a 3'-sialyl N-acetyllactosamine (3'-SLN) on a serine (S) residue. In the negative-ion mode, the CID MS(2) spectra of O-glycopeptides showed a relatively abundant glycoside-bond cleavage between the core N-acetylglucosamine (GlcNAc) and serine (S) that yields deprotonated C(3)-type fragment ions of O-glycan and deprotonated Z(0)-type peptide ions. The structure of the sLe(x) (3'-SLN) oligosaccharide was simply assigned by comparing the CID MS(3) spectrum derived from the C(3)-type fragment ion with the CID MS(2) spectra of the sLe(x) and sLe(a) (3'- and 6'-SLN) standards (i.e., negative-ion MS(n) spectral matching). The amino acid sequence of the peptide including the glycosylation site was determined from the ECD MS(2) spectrum in the positive-ion mode. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/17279605/Structural_analysis_of_O_glycopeptides_employing_negative__and_positive_ion_multi_stage_mass_spectra_obtained_by_collision_induced_and_electron_capture_dissociations_in_linear_ion_trap_time_of_flight_mass_spectrometry_ L2 - https://doi.org/10.1002/rcm.2885 DB - PRIME DP - Unbound Medicine ER -