Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry
Abstract
:1. Introduction
2. Results and Discussion
2.1. IMS MS Analysis of Hexasaccharide Domains in CS/DS from HEK293 DCN
2.2. IMS CID MS/MS for the Determination of Misregulations in the Sulfation Code
3. Materials and Methods
3.1. CS/DS Hexasaccharide Fraction
3.2. IMS MS and CID Tandem MS
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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No. | m/z exp | m/z Theor | Mass Accuracy (ppm) | Proposed Stucture | Sulfation | Molecular Ion |
---|---|---|---|---|---|---|
1 | 305.040 | 305.037 | −8.7 | 4,5-Δ-GlcAGalNAcIdoAGalNAc (2S) | 2S | [M-3H]3− |
2 | 314.309 | 314.306 | −11.1 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (1S) | 1S | [M-6H+2Na]4− |
3 | 323.308 | 323.304 | −13.2 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-4H]4− |
4 | 324.301 | 324.297 | −13.9 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (2S) | 2S | [M-5H+Na]4− |
5 | 343.296 | 343.293 | −8.7 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-4H]4− |
6 | 348.792 | 348.789 | −10.1 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-5H+Na]4− |
7 | 352.785 | 352.782 | −9.2 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-3H+K]2− |
8 | 354.281 | 354.284 | 8.5 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-6H+2Na]4− |
9 | 358.783 | 358.780 | −9.8 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (4S) | 4S | [M-4H]4− |
10 | 363.279 | 363.282 | 9.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-4H]4− |
11 | 372.774 | 372.771 | −8.1 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-5H+K]4− |
12 | 374.269 | 374.273 | 11.4 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-6H+2Na]4− |
13 | 384.990 | 384.994 | 11.3 | 4,5-Δ-GlcAGalNAcIdoAGalNAc (5S) | 5S | [M-3H]3− |
14 | 404.753 | 404.755 | 5.8 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (1S) | 1S | [M-3H]3− |
15 | 419.414 | 419.410 | −9.5 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (1S) | 1S | [M-5H+2Na]3− |
16 | 425.404 | 425.408 | 8.6 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (2S) | 2S | [M-3H]3− |
17 | 431.410 | 431.408 | −5.4 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-3H]3− |
18 | 437.411 | 437.408 | −7.6 | GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-3H]3− |
19 | 444.062 | 444.059 | −6.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-4H+K]3− |
20 | 449.064 | 449.060 | −8.9 | 4,5-Δ-GlcAGalNAcIdoAGalNAc - H2O (2S) | 2S | [M-2H]2− |
21 | 452.064 | 452.060 | −8.8 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (3S) | 3S | [M-3H]3− |
22 | 458.064 | 458.060 | −8.7 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-3H]3− |
23 | 465.390 | 465.387 | −5.7 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-4H+Na]3− |
24 | 470.714 | 470.712 | −5.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-4H+K]3− |
25 | 472.712 | 472.715 | 5.6 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-5H+2Na]3− |
26 | 478.035 | 478.039 | 8.4 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-5H+Na+K]3− |
27 | 480.046 | 480.042 | −8.3 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-6H+3Na]3− |
28 | 484.709 | 484.712 | 6.9 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-3H]3− |
29 | 492.041 | 492.040 | −2.7 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-4H+Na]3− |
30 | 497.365 | 497.364 | −2.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-4H+K]3− |
31 | 499.369 | 499.367 | −4.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-5H+2Na]3− |
32 | 510.014 | 510.016 | 3.9 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (4S) | 4S | [M-5H+2K]3− |
33 | 570.994 | 570.990 | −7.0 | 4,5-Δ-GlcAGalNAcIdoAGalNAc (4S) | 4S | [M-5H+3Na]2− |
34 | 629.617 | 629.619 | 3.2 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (1S) | 1S | [M-4H+2Na]2− |
35 | 638.612 | 638.610 | −3.1 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (2S) | 2S | [M-2H]2− |
36 | 647.620 | 647.616 | −7.0 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-2H]2− |
37 | 656.626 | 656.621 | −7.6 | GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-2H]2− |
38 | 678.595 | 678.589 | −8.8 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 - H2O (3S) | 3S | [M-2H]2− |
39 | 687.596 | 687.594 | −2.9 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-2H]2− |
40 | 698.584 | 698.585 | 1.4 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-3H+Na]2− |
41 | 706.569 | 706.572 | 4.2 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-3H+K]2− |
42 | 717.566 | 717.563 | −4.9 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-4H+Na+K]2− |
43 | 720.569 | 720.567 | −2.8 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-5H+3Na]2− |
44 | 899.133 | 899.128 | −5.6 | 4,5-Δ-GlcAGalNAcIdoAGalNAc - 2H2O (2S) | 2S | [M-2H]2− |
45 | 1296.242 | 1296.239 | −2.3 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (2S) | 2S | [M-H]− |
46 | 1376.198 | 1376.196 | −1.5 | 4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 (3S) | 3S | [M-H]− |
m/z | Charge State | Structure | The Type of Ion for the Structure in | |
---|---|---|---|---|
Figure 8a | Figure 8b | |||
96.964 | 1 | HOSO3 | ||
115.011 | 1 | GalNAc | 2,5X0 | |
157.027 | 1 | 4,5-Δ-GlcA | B1 | |
175.036 | 1 | 4,5-Δ-GlcA | C1 | |
193.050 | 1 | IdoA (0S) | Y2/Z1 or Y4/Z3 | |
199.006 | 1 | GalNAc (1S) | 0,2A6/B5 or0,2A4/B3 | |
237.544 | 1 | IdoA (1S) b | Z2/Y1 | Z4/Y3 |
282.045 | 1 | GalNAc (1S) a | Z1 | Z5/Z4 |
300.061 | 1 | GalNAc (1S) a | Y1 | Y5/Z4 |
307.559 | 2 | IdoAGalNAcIdoA (1S) a | Z4/Y1 | B3 |
343.318 | 1 | IdoAGalNAc (1S) a | 0,2X1 | 2,5A2 |
379.076 | 2 | GalNAcIdoAGalNAc (2S) a | Y3 | Z5/Z2 |
405.413 | 3 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | Y5 | |
418.068 | 3 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | Y5+K | |
432.071 | 3 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | 2,5A6 +K | |
440.079 | 1 | 4,5-Δ-GlcAGalNAc (1S) or IdoAGalNAc (1S) a | Z4/Y2 | B2 |
444.092 | 3 | GlcAGalNAcIdoAGalNAcIdoAGalNAc (3S) a | 1,5A6 | 1,5A6 |
454.023 | 1 | IdoAGalNAc (2S) b | 0,3X1 | 0,3X3/Z2 |
458.090 | 3 | GlcAGalNAcIdoAGalNAcIdoAGalNAc (3S) a | [M-3H]3− | |
468.062 | 2 | IdoAGalNAcIdoAGalNAc (2S) a | Y4 | |
470.714 | 3 | 4,5-Δ-GlcAGalNAcIdoAGalNAcIdoAGalNAc (3S) a | [M-4H+K]3− | |
476.100 | 1 | IdoAGalNAc (1S) a | Y4/Z2 | Y3/Z1 orY4/Z2 |
498.077 | 2 | IdoAGalNAcIdoAGalNAc (3S) b | Z4 | C4 |
507.085 | 2 | IdoAGalNAcIdoAGalNAc (3S) b | Y4 | Y5/Z1 |
538.054 | 1 | 4,5-Δ-GlcAGalNAcIdoA (0S) | B3 | |
IdoAGalNAc (2S) b | Z2 | Y4/Y2 | ||
543.091 | 2 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | 1,4X4 | |
556.085 | 2 | 4,5-Δ-GlcAGalNAcIdoAGalNAcIdoA (2S) a | B5 +K | |
565.092 | 2 | 4,5-Δ-GlcAGalNAcIdoAGalNAcIdoA (2S) a | C5 +K | |
568.065 | 2 | GalNAcIdoAGalNAcIdoAGalNAc (2S) | Y5 | |
608.625 | 2 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | Y5 | |
612.035 | 1 | IdoAGalNAcIdoA (2S) b | 1,4A5/C2 | 1,4A3 |
616.126 | 1 | 4,5-Δ-GlcAGalNAcIdoA (1S) a IdoAGalNAcIdoA (1S) a | - Z4/Y1 | B3 Z4/Y1 |
622.627 | 2 | 4,5-Δ-GlcAGalNAcIdoAGalNAcIdoAGalNAc (3S) a | 1,5X5 | |
627.595 | 2 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | Y5+K | |
630.071 | 2 | 4,5-Δ-GlcAGalNAcIdoAGalNAcIdoAGalNAc (3S) a | 2,5A6 | |
648.613 | 2 | GalNAcIdoAGalNAcIdoAGalNAc (3S) a | 2,5A6 +K | |
654.075 | 1 | IdoAGalNAcIdoA (1S) a | Z4/Y1+K | B3+K |
779.097 | 1 | GalNAcIdoAGalNAc (3S) b | 2,4X2 | 2,4X4/Y2 |
797.109 | 1 | GalNAcIdoAGalNAc (2S) a | Y3+K | Y5/Z2+K |
821.112 | 1 | GalNAcIdoAGalNAc (3S) b | Z3 | Y5/Y2 |
839.132 | 1 | GalNAcIdoAGalNAc (3S) b | Y3 | Y5/Z2 |
857.169 | 1 | 4,5-Δ-GlcAGalNAcIdoAGalNAc (1S) | B4+K | B4+K or Y5/Y1+K |
875.179 | 1 | 4,5-Δ-GlcAGalNAcIdoAGalNAc (1S) | C4+K | C4+K or Y5/Z1+K |
911.152 | 1 | IdoAGalNAcIdoAGalNAc (3S) b | 0,3X3 | C4/0,3A1 |
917.187 | 1 | IdoAGalNAcIdoAGalNAc (2S) a | Z4 | Z4 |
955.139 | 1 | IdoAGalNAcIdoAGalNAc (3S) b IdoAGalNAcIdoAGalNAc (2S) a | 2,4X3 Z4+K | 2,4X4/Z1 Z4+K |
997.154 | 1 | IdoAGalNAcIdoAGalNAc (3S) b | Z4 | Y5/Y1 |
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Sarbu, M.; Ica, R.; Sharon, E.; Clemmer, D.E.; Zamfir, A.D. Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry. Molecules 2022, 27, 6026. https://doi.org/10.3390/molecules27186026
Sarbu M, Ica R, Sharon E, Clemmer DE, Zamfir AD. Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry. Molecules. 2022; 27(18):6026. https://doi.org/10.3390/molecules27186026
Chicago/Turabian StyleSarbu, Mirela, Raluca Ica, Edie Sharon, David E. Clemmer, and Alina D. Zamfir. 2022. "Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry" Molecules 27, no. 18: 6026. https://doi.org/10.3390/molecules27186026