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. 2013 Jul;24(7):1072-81.
doi: 10.1007/s13361-013-0624-y. Epub 2013 Apr 30.

Unusual fragmentation pathways in collagen glycopeptides

Irina Perdivara  1 , Lalith Perera, Marnisa Sricholpech, Masahiko Terajima, Nancy Pleshko, Mitsuo Yamauchi, Kenneth B Tomer
Affiliations

Unusual fragmentation pathways in collagen glycopeptides

Irina Perdivara et al. J Am Soc Mass Spectrom. 2013 Jul.

Abstract

Collagens are the most abundant glycoproteins in the body. One characteristic of this protein family is that the amino acid sequence consists of repeats of three amino acids -(X-Y-Gly)n. Within this motif, the Y residue is often 4-hydroxyproline (HyP) or 5-hydroxylysine (HyK). Glycosylation in collagen occurs at the 5-OH group in HyK in the form of two glycosides, galactosylhydroxylysine (Gal-HyK) and glucosyl galactosylhydroxylysine (GlcGal-HyK). In collision induced dissociation (CID), collagen tryptic glycopeptides exhibit unexpected gas-phase dissociation behavior compared to typical N- and O-linked glycopeptides (i.e., in addition to glycosidic bond cleavages, extensive cleavages of the amide bonds are observed). The Gal- or GlcGal- glycan modifications are largely retained on the fragment ions. These features enable unambiguous determination of the amino acid sequence of collagen glycopeptides and the location of the glycosylation site. This dissociation pattern was consistent for all analyzed collagen glycopeptides, regardless of their length or amino acid composition, collagen type or tissue. The two fragmentation pathways-amide bond and glycosidic bond cleavage-are highly competitive in collagen tryptic glycopeptides. The number of ionizing protons relative to the number of basic sites (i.e., Arg, Lys, HyK, and N-terminus) is a major driving force of the fragmentation. We present here our experimental results and employ quantum mechanics calculations to understand the factors enhancing the labile character of the amide bonds and the stability of hydroxylysine glycosides in gas phase dissociation of collagen glycopeptides.

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Figures

Figure 1
Figure 1
Ion trap CID of glycopeptides α-1 [76–90], for the precursor ions: A. m/z 558.7 (3+), B. m/z 612.7 (3+), and C. m/z 918.6 (2+). Each glycopeptide contains oxidized Met (Mox), two HyP and either Gal-HyK or GlcGal-HyK, as indicated. Arrows indicate loss of CH3SOH (−64 Da) from Mox.
Figure 2
Figure 2
Deconvoluted QTof CID spectra of the 3+ and 4+ precursor ions of three glycopeptides: A. and B. – bovine type II collagen α-1 [193–237]; C. and D. – bovine type II collagen α-1 [91–126]; E. and F. – bovine type I collagen α-1 [193–237]. Following collision energy ramps were employed: 20 – 30 V for the spectra in A., B. and D., and 30 – 40 V for the spectra in C., E. and F.
Figure 3
Figure 3
Deconvoluted QTof CID spectra of glycopeptide ions of: A. m/z 851.143 (4+), corresponding to α-1 [556–585], and B. m/z 659.663 (3+), corresponding to α-1 [556–573] from mouse type I collagen. The spectra were obtained with a collision energy ramp of 30 – 40 V (A.), and 25 – 30 V (B.).
Figure 4
Figure 4
A. One lowest energy structure of H3N+-Gly-Gly-HyP-NMe determined at the wB97-XD level of Gaussian calculations with the 6-311++G** basis set; B. Energy profile for the proton transfer from protonated N-terminus to the second amide nitrogen in Gly-Gly-HyP; C. Lowest energy structure determined for the system βCH3-γCH2-δCH(O-β-D-Gal)-εCH2-N+H3 when the ionizing proton is at the ε-amino group, and D. when the ionizing proton was transferred to the glycosidic oxygen.
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References

    1. Varki A, Cummings R, Esko J, Freeze H, Hart G, Marth J. Essentials of Glycobiology. 2009 - PubMed
    1. Settineri CA, Medzihradszky KF, Masiarz FR, Burlingame AL, Chu C, George-Nascimento C. Characterization of O-glycosylation sites in recombinant B-chain of platelet-derived growth factor expressed in yeast using liquid secondary ion mass spectrometry, tandem mass spectrometry and Edman sequence analysis. Biomed Environ Mass Spectrom. 1990;19(11):665–76. - PubMed
    1. Medzihradszky KF, Gillece-Castro BL, Hardy MR, Townsend RR, Burlingame AL. Structural Elucidation of O-linked Glycopeptides by High Energy Collision-Induced Dissociation. Journal of the American Society for Mass Spectrometry. 1996;7:319–328. - PubMed
    1. Hanisch FG, Green BN, Bateman R, Peter-Katalinic J. Localization of O-glycosylation sites of MUC1 tandem repeats by QTOF ESI mass spectrometry. J Mass Spectrom. 1998;33(4):358–62. - PubMed
    1. Alving K, Paulsen H, Peter-Katalinic J. Characterization of O-glycosylation sites in MUC2 glycopeptides by nanoelectrospray QTOF mass spectrometry. J Mass Spectrom. 1999;34(4):395–407. - PubMed

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