Table 1. Detected Glycosylated Products Elongated on Xyl-Ser-C12a.
| structure | ion type | theoretical m/z | experimental m/z | error (ppm) | retention time (min) | proportion (%) | fragment ions |
|---|---|---|---|---|---|---|---|
| Hex-Xyl-R | [M – H]− | 579.3134 | 579.3116 | –3.11 | 6.90 | 7.41 | 267.2083 (Z0), 161.0441 (B1) |
| Hex(S)-Xyl-R | [M – H]− | 659.2703 | 659.2704 | 0.15 | 8.66 | 0.44 | 391.0555 (C2), 241.0023 (B1), 96.9610 ([HSO4]−) |
| Hex-Hex-Xyl-R | [M – H]− | 741.3663 | 741.3636 | –3.64 | 11.24 | 11.86 | 395.1242 (2,5A3), 383.1205 (2,4A3), 323.0990 (B2), 267.2075 (Z0), 263.0779 (2,5A2), 221.0665 (2,4A2) |
| Hex(S)-Hex-Xyl-R | [M – H]− | 821.3231 | 821.3158 | –8.89 | 11.99 | 1.28 | 553.1039 (C3), 403.0630 (B2), 241.0020 (B1), 96.9598 ([HSO4]−) |
| Hex-Hex(S)-Xyl-R | [M – H]− | 821.3231 | 821.3193 | –4.63 | 12.55 | 0.72 | 659.2714 (Y2), 553.1146 (C3), 403.0518 (B2), 96.9564 ([HSO4]−) |
| Hex-Xyl(P)-R | [M – H]− | 659.2798 | 659.2767 | –4.70 | 13.34 | 17.85 | 497.2288 (Y1), 391.0667 (C2), 373.0561 (B1), 259.0222 ([Hex + PO3]−), 241.0112 ([Hex – H2O + PO3]−), 78.9585 ([PO3]−) |
| NeuAc-Hex-Xyl-R | [M – H]− | 870.4089 | 870.4049 | –4.60 | 13.41 | 10.24 | 649.3218 (0,2X3), 602.1959 (C3), 480.1747 (2,5A3 – CO2), 468.1740 (2,4A3 – CO2), 408.1616 (B3 – CO2), 290.0983 (B1), 267.2126 (Z0) |
| Hex-Hex-Xyl(P)-R | [M – H]− | 821.3326 | 821.3297 | –3.53 | 16.06 | 6.22 | 659.2807 (Y2), 553.1211 (C3), 535.1099 (B3), 497.2257 (Y1), 421.0768 ([Hex + Hex + PO3]−), 78.9583 ([PO3]−) |
| NeuAc-Hex-Hex-Xyl-R_1 | [M – H]− | 1032.4617 | 1032.4546 | –6.88 | 16.35 | 0.13 | 811.3792 (0,2X3), 642.2279 (2,5A3 – CO2), 570.2025 (B2 – CO2), 395.1175 (2,5A3/Y3), 383.1174 (2,4A3/Y3), 323.0972 (B2/Y3), 290.0881 (B1), 267.2068 (Z0) |
| Hex(S)-Hex-Xyl(P)-R | [M – H]− | 901.2894 | 901.2847 | –5.21 | 17.04 | 0.07 | 821.3353 ([M – H – SO3]−), 659.2846 (Y2), 553.1113 (C3 – SO3), 535.1119 (B3 – SO3), 240.9993 (B1), 78.9557 ([PO3]−) |
| NeuAc-Hex-Xyl(P)-R_1 | [M – H]− | 950.3752 | 950.376 | 0.84 | 17.96 | 5.81 | 906.3903 ([M – H – CO2]−), 659.2913 (Y2), 497.2286 (Y1), 391.0692 (C3/Y2), 373.0564 (B3/Y2), 78.9587 ([PO3]−) |
| HexA-Hex-Hex-Xyl-R | [M – H]− | 917.3978 | 917.398 | 0.22 | 18.17 | 1.35 | 649.1868 (C4), 589.1686 (0,2A4), 517.1450 (C3), 455.1406 (B4/Y3), 395.1209 (2,5A4/Y3), 337.0771 (B2), 323.1008 (B3/Y3), 267.2066 (Z0) |
| HexA(S)-Hex-Hex-Xyl-R | [M – H]− | 997.3552 | 997.3514 | –3.81 | 18.35 | 1.32 | 917.4081 ([M – H – SO3]−), 729.1453 (C4), 649.1871 (C4 – SO3), 267.2077 (Z0), 254.9880 (B1), 210.9911 (B1 – CO2), 96.9594 ([HSO4]−) |
| NeuAc-NeuAc-Hex-Xyl-R | [M – H]− | 1161.5043 | 1161.4995 | –4.13 | 18.28 | 0.05 | 870.4119 (Y3), 581.1873 (B2), 537.1964 (B2 – CO2), 290.0938 (B1), 267.2074 (Z0) |
| HexNAc-HexA-Hex-Hex-Xyl-R_1 | [M – H]− | 1120.4777 | 1120.4736 | –3.66 | 19.52 | 0.09 | 899.3929 (Z4), 509.1490 (2,5A5/Z4 – CO2), 437.1272 (C4/Z4 – CO2), 267.2029 (Z0) |
| HexNAc-HexA-Hex-Hex-Xyl-R_2 | [M – H]− | 1120.4777 | 1120.4736 | –3.66 | 19.69 | 10.62 | 899.3914 (Z4), 855.4024 (Z4 – CO2), 852.2647 (C5), 509.1566 (2,5A5/Z4 – CO2), 437.1315 (C4/Z4 – CO2), 267.2079 (Z0) |
| HexNAc(S)-HexA-Hex-Hex-Xyl-R_1 | [M – 2H]2– | 599.7148 | 599.7127 | –3.50 | 19.96 | 0.09 | 932.2285 (C5), 852.2615 (C5 – SO3), 800.1821 (C4), 720.2226 (C4 – SO3), 638.1266 (C3), 558.1722 (C3 – SO3), 476.0723 (C2), 458.0573 (B2), 396.1162 (C2 – SO3), 300.0408 (C1), 267.2064 (Z0) |
| NeuAc-Hex-Hex-Xyl(P)-R | [M – H]− | 1112.428 | 1112.4242 | –3.42 | 20.13 | 0.31 | 1068.4458 ([M – H – CO2]−), 821.3354 (Y3), 659.2799 (Y2), 553.1210 (C4/Y3), 535.1100 (B4/Y3), 497.2289 (Y1), 290.0853 (B1), 79.9585 ([PO3]−) |
| HexA-Hex-Hex-Xyl(P)-R | [M – H]− | 997.3647 | 997.3585 | –6.22 | 21.72 | 0.30 | 821.3367 (Y3), 729.1527 (C4), 659.2758 (Y2), 553.1232 (C4/Y2), 535.1155 (B4/Y2), 497.2288 (Y1), 78.9577 ([PO3]−) |
| HexA(S)-Hex-Hex-Xyl(P)-R | [M – 2H]2– | 538.1578 | 538.1541 | –6.88 | 21.96 | 0.37 | 821.3328 (Y3), 254.9837 (B1), 78.9596 ([PO3]−) |
| HexNAc-HexA-Hex-Hex-Xyl(P)-R | [M – 2H]2– | 599.7184 | 599.7173 | –1.83 | 22.83 | 20.91 | 979.3587 (Z4), 935.3710 (Z4 – CO2), 821.3344 (Y3), 803.3246 (Z3), 659.2821 (Y2), 553.1206 (C5/Y3), 535.1094 (B5/Y3), 497.2276 (Y1), 355.0439 (B5/Z2), 78.9583 ([PO3]−) |
| NeuAc-NeuAc-Hex-Hex-Xyl(P)-R_1 | [M – 2H]2– | 701.2581 | 701.2564 | –2.42 | 22.86 | 0.63 | 821.3374 (Y2), 290.0917 (B1), 78.9576 ([PO3]−) |
| NeuAc-NeuAc-Hex-Hex-Xyl(P)-R_2 | [M – 2H]2– | 701.2581 | 701.2554 | –3.85 | 23.41 | 0.03 | 821.3371 (Y2), 290.0862 (B1), 78.9579 ([PO3]−) |
| HexA-HexNAc-HexA-Hex-Hex-Xyl-R | [M – 2H]2– | 647.7513 | 647.7484 | –4.48 | 24.39 | 0.05 | 1028.2963 (C6), 896.2433 (C5), 734.1983 (C4), 572.1512 (C3), 509.1527 (2,5A6/Z4 – CO2), 396.1161 (C2), 267.2080 (Z0), 193.0338 (C1) |
| (HexNAc-HexA)2-Hex-Hex-Xyl-R_HS | [M – 2H]2– | 749.291 | 749.2877 | –4.40 | 24.45 | 0.03 | 1234.4941 (Z6 – CO2), 899.4190 (Z4), 757.2145 (B4), 599.1989 (C3), 509.1496 (2,5A7/Z4 – CO2), 480.1401 (2,5A3), 396.1109 (C2), 267.2070 (Z0) |
| (HexNAc-HexA)2-Hex-Hex-Xyl-R_CS | [M – 2H]2– | 749.291 | 749.2887 | –3.07 | 24.97 | 0.91 | 1234.5146 (Z6 – CO2), 1231.3741 (C7), 1010.2892 (C7/Z6), 966.3002 (C7/Z6 – CO2), 899.3911 (Z4), 757.2137 (B4), 599.1954 (C3), 509.1534 (2,5A7/Z4 – CO2), 396.1157 (C2), 267.2076 (Z0) |
| HexNAc(S)-HexA-HexNAc-HexA-Hex-Hex-Xyl-R | [M – 2H]2– | 789.2694 | 789.2659 | –4.43 | 25.21 | 0.36 | 1120.4812 (Y5), 1010.2779 (C7/Z6), 917.4000 (Y4), 899.3916 (Z4), 757.2137 (B4 – SO3), 679.1526 (C3), 599.1962 (C3 – SO3), 509.1509 (2,5A7/Z4 – CO2), 458.0632 (B2), 396.1171 (C2 – SO3), 300.0399 (C1), 282.0290 (B1), 267.2072 (Z0) |
| (HexNAc-HexA)2-Hex-Hex-Xyl(P)-R_2 | [M – 2H]2– | 789.2741 | 789.2722 | –2.41 | 27.31 | 0.49 | 1234.5221 (Z6 – CO2 – HPO3), 997.3666 (Y4), 979.3548 (Z4), 935.3652 (Z6 – CO2), 821.3378 (Y3), 659.2739 (Y2), 396.1106 (C2), 78.9588 ([PO3]−) |
| (HexNAc-HexA)3-Hex-Hex-Xyl-R | [M – 2H]2– | 938.8467 | 938.8445 | –2.34 | 28.90 | 0.05 | 1278.5253 (Z6), 1120.4763 (Y5), 978.3136 (C5), 917.4018 (Y4), 899.3984 (Z4), 775.2314 (C4), 757.2167 (B4), 599.1964 (C3), 509.1504 (2,5A9/Z4 – CO2), 396.1187 (C2), 267.2078 (Z0) |
a
Proportions were calculated from the peak areas of the glycosylated products.