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Direct structural assignment of neutral and sialylated N-glycans of glycopeptides using collision-induced dissociation MSn spectral matching.
Rapid Commun Mass Spectrom. 2006; 20(23):3557-65.RC

Abstract

Mass spectrometric analyses of various N-glycans binding to proteins and peptides are highly desirable for elucidating their biological roles. An approach based on collision-induced dissociation (CID) MS(n) spectra acquired by electrospray ionization linear ion trap time-of-flight mass spectrometry (ESI-LIT-TOFMS) in the positive- and negative-ion modes has been proposed as a direct method of assigning N-glycans without releasing them from N-glycopeptides. In the positive-ion mode of this approach, the MS(2) spectrum of N-glycopeptide was acquired so that a glycoside-bond cleavage occurs in the chitobiose residue (i.e., GlcNAcbeta1-4GlcNAc, GlcNAc: N-acetylglucosamine) attached to asparagine (N), and two charges on the [M+H+Na](2+) precursor ion are shared with both of the resulting fragments. These fragments are sodiated B(n)-type fragment ions of oligosaccharide (N-glycan) and a protonated peptide ion retaining one GlcNAc residue on the asparagine (N) residue. The structure of N-glycan was assigned by comparing MS(3) spectra derived from both the sodiated B(n)-type fragment ions of N-glycopeptide and the PA (2-aminopyridine) N-glycan standard (i.e., MS(n) spectral matching). In a similar manner, the structural assignment of sialylated N-glycan was performed by employing the negative-ion CID MS(n) spectra of deprotonated B(n)-type fragment ions of N-glycopeptide and the PA N-glycan standard. The efficacy of this approach was tested with chicken egg yolk glycopeptides with a neutral and a sialylated N-glycan, and human serum IgG glycopeptides with neutral N-glycan isomers. These results suggest that the approach based on MS(n) spectral matching is useful for the direct and simple structural assignment of neutral and sialylated N-glycans of glycopeptides.

Authors+Show Affiliations

Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.No 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

17091533

Citation

Ito, Hiroki, et al. "Direct Structural Assignment of Neutral and Sialylated N-glycans of Glycopeptides Using Collision-induced Dissociation MSn Spectral Matching." Rapid Communications in Mass Spectrometry : RCM, vol. 20, no. 23, 2006, pp. 3557-65.
Ito H, Takegawa Y, Deguchi K, et al. Direct structural assignment of neutral and sialylated N-glycans of glycopeptides using collision-induced dissociation MSn spectral matching. Rapid Commun Mass Spectrom. 2006;20(23):3557-65.
Ito, H., Takegawa, Y., Deguchi, K., Nagai, S., Nakagawa, H., Shinohara, Y., & Nishimura, S. (2006). Direct structural assignment of neutral and sialylated N-glycans of glycopeptides using collision-induced dissociation MSn spectral matching. Rapid Communications in Mass Spectrometry : RCM, 20(23), 3557-65.
Ito H, et al. Direct Structural Assignment of Neutral and Sialylated N-glycans of Glycopeptides Using Collision-induced Dissociation MSn Spectral Matching. Rapid Commun Mass Spectrom. 2006;20(23):3557-65. PubMed PMID: 17091533.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Direct structural assignment of neutral and sialylated N-glycans of glycopeptides using collision-induced dissociation MSn spectral matching. AU - Ito,Hiroki, AU - Takegawa,Yasuhiro, AU - Deguchi,Kisaburo, AU - Nagai,Shinji, AU - Nakagawa,Hiroaki, AU - Shinohara,Yasuro, AU - Nishimura,Shin-Ichiro, PY - 2006/11/9/pubmed PY - 2007/1/12/medline PY - 2006/11/9/entrez SP - 3557 EP - 65 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 20 IS - 23 N2 - Mass spectrometric analyses of various N-glycans binding to proteins and peptides are highly desirable for elucidating their biological roles. An approach based on collision-induced dissociation (CID) MS(n) spectra acquired by electrospray ionization linear ion trap time-of-flight mass spectrometry (ESI-LIT-TOFMS) in the positive- and negative-ion modes has been proposed as a direct method of assigning N-glycans without releasing them from N-glycopeptides. In the positive-ion mode of this approach, the MS(2) spectrum of N-glycopeptide was acquired so that a glycoside-bond cleavage occurs in the chitobiose residue (i.e., GlcNAcbeta1-4GlcNAc, GlcNAc: N-acetylglucosamine) attached to asparagine (N), and two charges on the [M+H+Na](2+) precursor ion are shared with both of the resulting fragments. These fragments are sodiated B(n)-type fragment ions of oligosaccharide (N-glycan) and a protonated peptide ion retaining one GlcNAc residue on the asparagine (N) residue. The structure of N-glycan was assigned by comparing MS(3) spectra derived from both the sodiated B(n)-type fragment ions of N-glycopeptide and the PA (2-aminopyridine) N-glycan standard (i.e., MS(n) spectral matching). In a similar manner, the structural assignment of sialylated N-glycan was performed by employing the negative-ion CID MS(n) spectra of deprotonated B(n)-type fragment ions of N-glycopeptide and the PA N-glycan standard. The efficacy of this approach was tested with chicken egg yolk glycopeptides with a neutral and a sialylated N-glycan, and human serum IgG glycopeptides with neutral N-glycan isomers. These results suggest that the approach based on MS(n) spectral matching is useful for the direct and simple structural assignment of neutral and sialylated N-glycans of glycopeptides. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/17091533/Direct_structural_assignment_of_neutral_and_sialylated_N_glycans_of_glycopeptides_using_collision_induced_dissociation_MSn_spectral_matching_ L2 - https://doi.org/10.1002/rcm.2761 DB - PRIME DP - Unbound Medicine ER -