Comprehensive Detection of Isopeptides between Human Tissue Transglutaminase and Gluten Peptides
Abstract
:1. Introduction
2. Materials and Methods
2.1. Material
2.2. Enzyme Activity Test of TG2
2.3. Isopeptide Standard
2.4. Model Reaction of TG2 and PepQ
2.5. Model Reaction at Different Molar Ratios
2.6. Model Reaction with Three Different Model Peptides
2.7. Tryptic Digestion and Clean-Up by Solid Phase Extraction
2.8. Nanoscale Liquid Chromatography-Tandem Mass Spectrometry
2.9. Isopeptide Identification Using MaxQuant
2.10. Assignment of MS/MS Fragments of the Isopeptide Sequences Using ProteinProspector
2.11. Isopeptide Confirmation Using Skyline
2.12. Isopeptide Identification Using pLink
2.13. 3D-Structure Model of TG2
3. Results
3.1. Determination of TG2 Enzyme Activity
3.2. Identification of the Isopeptide Standard
3.3. Identification of TG2-Peptides Involved in Isopeptide Formation
3.4. Identification of TG2-TG2 Crosslinks
3.5. Identification of Crosslinks and Deamidation Sites Within PepQ
3.6. Visualization of Isopeptides with Skyline
3.7. Estimation of Preferred Lysine Residues
3.8. Location of the Complex-Forming Lysine Residues in the 3D-Model of TG2
3.9. Identification of Isopeptides in the Extended Model System
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Data Availability
References
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(A) TG2-PepQ Isopeptides with Unambiguous Modification Sites (Localization Probability >75% for the Binding Site in PepQ) | ||||||||||
Lysine Residue a | Sequence of TG2 peptide | Modification | Precursor | Charge State | MaxQuant Score | No. of Fragment Matches | Q6 Isopeptide Probabilities | Identification E-value pLink2 b | ||
β-side c | α-side d,e | m/z | α f | β g | α | β | [%] | |||
173 | QEYVLTQQGFIYQGSAKFIK | PFPQPQLPY | 1139.59 | 3+ | 30.17 | 128.14 | 6 | 32 | 4.2 | 4.40 × 10−4 |
205 | FLKNAGR | PFPQPQLPY | 625.01 | 3+ | 135.77 | 35.63 | 18 | 10 | 100 | 2.24 × 10−11 |
PFPEPQLPY | 625.34 | 3+ | 117.16 | 19.92 | 15 | 8 | 99.7 | - | ||
265 | WKNHGCQR | PFPQPQLPY | 699.36 | 3+ | 79.45 | 29.81 h | 12 | 4 | 100 | 6.36 × 10−6 |
380 | AIKEGDLSTK | PFPQPQLPY | 1065.07 | 2+ | 151.17 | 29.74 | 17 | 7 | 100 | 6.56 × 10−7 |
PFPQPELPY | 710.71 | 3+ | 94.14 | 45.88 | 14 | 14 | 1.7 | - | ||
425 | SLIVGLKISTK | PFPQPQLPY | 742.77 | 3+ | 79.94 | 84.48 | 12 | 21 | 98.0 | 7.57 × 10−6 |
429 | ISTKSVGR | PFPQPQLPY | 639.02 | 3+ | 107.90 | 26.96 h | 17 | 5 | 100 | 4.57 × 10−6 |
429 | ISTKSVGRDER | PFPQPQLPY | 579.56 | 4+ | 168.23 | - i | 22 | - h | 100 | - |
444 | EDITHTYKYPEGSSEER j | PFPQPQLPY | 1036.82 | 3+ | 133.60 | 141.60 | 17 | 33 | 100 | 2.74 × 10−8 |
PFPQPELPY | 1037.15 | 3+ | 119.29 | 96.45 | 14 | 29 | 0.3 | - | ||
444 | DEREDITHTYKYPEGSSEER | PFPQPQLPY | 877.91 | 4+ | 63.57 | 53.59 | 9 | 17 | 89.7 | - |
464 | ANHLNKLAEK | PFPQPQLPY | 735.73 | 3+ | 155.82 | 47.98 | 22 | 16 | 100 | - |
464 | ANHLNKLAEKEETGMAMR | PFPQPQLPY | 622.92 | 5+ | 110.31 | 92.31 | 17 | 25 | 100 | 1.07 × 10−6 |
PFPEPQLPY | 623.11 | 5+ | 121.99 | 78.02 | 14 | 22 | 0.2 | - | ||
468 | LAEKEETGMAMR | PFPQPQLPY | 811.73 | 3+ | 116.73 | 95.09 | 17 | 21 | 100 | 6.50 × 10−11 |
PFPEPQLPY | 812.06 | 3+ | 73.93 | 38.77 | 12 | 12 | 99.5 | - | ||
562 | DCLTESNLIKVR j | PFPQPQLPY | 820.10 | 3+ | 92.47 | 199.82 | 14 | 31 | 100 | 3.01 × 10−8 |
562 | YRDCLTESNLIKVR | PFPQPQLPY | 695.12 | 4+ | 157.86 | 111.74 | 19 | 26 | 100 | - |
590 | DLYLENPEIKIR j | PFPQPQLPY | 1285.67 | 2+ | 164.89 | 183.03 | 19 | 28 | 100 | 2.62 × 10−6 |
PFPQPELPY | 1286.18 | 2+ | 59.21 | 108.56 | 8 | 21 | 13 | - | ||
598 | ILGEPKQK | PFPQPQLPY | 660.73 | 3+ | 118.01 | 57.18 h | 14 | 10 | 99.9 | 5.38 × 10−7 |
598 | ILGEPKQKR | PFPQPQLPY | 712.74 | 3+ | 108.64 | 51.95 | 15 | 12 | 100 | 3.56 × 10−7 |
600 | QKR k | PFPQPQLPY | 749.91 | 2+ | 195.98 | - i | 20 | -h | 100 | - |
649 | TVEIPDPVEAGEEVKVR i | PFPQPQLPY | 978.85 | 3+ | 29.16 | 122.10 | 10 | 24 | - | 1.04 × 10−2 |
663 | MDLLPLHMGLHKLVVNFESDKLK | PFPQPQLPY | 937.01 | 4+ | 93.20 | 103.02 | 15 | 32 | 98.5 | - |
677 | AVKGFR j | PFPQPQLPY | 872.98 | 2+ | 168.74 | 22.29 g | 19 | 7 | 100 | 1.04 × 10−6 |
PFPQPELPY | 873.47 | 2+ | 151.51 | - i | 17 | -h | 0 | - | ||
(B) TG2-PepQ Isopeptides with Ambiguous Modification Sites | ||||||||||
464 | ANHLNKLAEK | PFPEPQLPY | 552.30 | 4 | 53.17 | 48.28 | 9 | 14 | 50 | - |
550 | SVPLCILYEKYR | PFPQPQLPY | 851.12 | 3 | - i | 47.92 | - i | 11 | - | - |
598/600 | ILGEPKQK l | PFPQPELPY | 661.03 | 3+ | 98.30 | 65.25 | 12 | 10 | 6.8 | - |
600/602 | QKRKj k,l | PFPQPQLPY | 542.98 | 3+ | 121.99 | - i | 15 | - h | 100 | - |
672/674 | LVVNFESDKLK | PFPQPQLPY | 787.09 | 3+ | 104.75 | 103.91 | 17 | 20 | 100 | 8.51 × 10−7 |
672/674 | LVVNFESDKLKAVK h,l | PFPQPQLPY | 665.12 | 4+ | 50.70 | 52.27 | 9 | 13 | 97.2 | 1.57 × 10−2 b |
Sequence of Isopeptide a | Precursor | Charge State | Residues in TG2 b | E-value pLink2 c | Number of MS2 Scans d | |
---|---|---|---|---|---|---|
m/z | K | Q | ||||
VVSGMVNCNDDQGVLLGR/EKLVVR | 2601.35 | 4+ | 30 | 234 | 2.46 × 10−12 | 6 |
VVSGMVNCNDDQGVLLGR/FLKNAGR | 2663.34 | 3+ | 205 | 234 | 1.77 × 10−17 | 10 |
QEYVLTQQGFIYQGSAK/AIKEGDLSTK | 3003.53 | 3+ | 380 | 164 | 3.48 × 10−15 | 3 |
VVSGMVNCNDDQGVLLGR/AIKEGDLSTK | 2919.45 | 3+ | 380 | 234 | 1.96 × 10−15 | 5 |
AIKEGDLSTK/NHGCQR | 1757.86 | 4+ | 380 | 270 | 1.50 × 10−4 | 6 |
NEFGEIQGDK/AIKEGDLSTK | 2180.07 | 3+ | 380 | 324 | 7.94 × 10−7 | 2 |
VVTNYNSAHDQNSNLLIEYFR/AIKEGDLSTK e | 3540.76 | 4+ | 380 | 307 | 2.23 × 10−7 | 6 |
VVSGMVNCNDDQGVLLGR/ISTKSVGRDER | 3105.54 | 5+ | 429 | 234 | 1.07 × 10−3 | 3 |
EDITHTYKYPEGSSEER/VVSGMVNCNDDQGVLLGR | 3898.79 | 5+ | 444 | 234 | 1.62 × 10−8 | 6 |
EDITHTYKYPEGSSEER/WKNHGCQR | 3051.36 | 5+ | 444 | 270 | 4.24 × 10−3 | 2 |
VVTNYNSAHDQNSNLLIEYFR/EDITHTYKYPEGSSEER | 4520.09 | 6+ | 444 | 307 | 3.36 × 10−12 | 7 |
EDITHTYKYPEGSSEER/NEFGEIQGDK | 3159.40 | 5+ | 444 | 324 | 1.94 × 10−3 | 4 |
VGQSMNMGSDFDVFAHITNNTAEEYVCR/EDITHTYKYPEGSSEER e | 5158.24 | 5+ | 444 | 481 | 3.54 × 10−11 | 3 |
VVSGMVNCNDDQGVLLGR/ANHLNKLAEK | 2995.51 | 4+ | 464 | 234 | 2.93 × 10−9 | 4 |
VVTNYNSAHDQNSNLLIEYFR/ANHLNKLAEK e | 3616.81 | 5+ | 464 | 307 | 1.20 × 10−6 | 10 |
VGQSMNMGSDFDVFAHITNNTAEEYVCR/ANHLNKLAEK | 4254.96 | 5+ | 464 | 481 | 9.92 × 10−8 | 5 |
VGQSMNMGSDFDVFAHITNNTAEEYVCR/LEAKEETGMAMR e | 4482.98 | 5+ | 468 | 481 | 1.20 × 10−17 | 4 |
QEYVLTQQGFIYQGSAK/DLYLENPEIKIR | 3444.77 | 4+ | 590 | 164 | 6.77 × 10−5 | 3 |
VVSGMVNCNDDQGVLLGR/DLYLENPEIKIR | 3360.69 | 4+ | 590 | 234 | 7.07 × 10−11 | 7 |
DLYLENPEIKIR/WKNHGCQR | 2513.27 | 4+ | 590 | 270 | 2.01 × 10−11 | 5 |
VVTNYNSAHDQNSNLLIEYFR/DLYLENPEIKIR | 3981.99 | 5+ | 590 | 307 | 3.04 × 10−7 | 4 |
ILGEPKQK/AVKGFR | 1571.92 | 4+ | 599 | 677 | 1.03 × 10−10 | 4 |
ILGEPKQKR/AVKGFR | 1728.03 | 4+ | 599 | 677 | 1.21 × 10−3 | 3 |
VVSGMVNCNDDQGVLLGR/TVEIPDPVEAGEEVKVR e | 3724.85 | 4+ | 649 | 234 | 6.78 × 10−13 | 6 |
TVEIPDPVEAGEEVKVR/NHGCQR | 2563.26 | 4+ | 649 | 270 | 5.69 × 10−3 | 2 |
VVTNYNSAHDQNSNLLIEYFR/TVEIPDPVEAGEEVKVR | 4346.16 | 5+ | 649 | 307 | 7.66 × 10−5 | 4 |
LVVNFESDKLKAVK/NHGCQR | 2286.20 | 5+ | 672 | 270 | 8.80 × 10−6 | 2 |
QEYVLTQQGFIYQGSAK/AVKGFR | 2619.36 | 4+ | 677 | 164 | 5.82 × 10−9 | 6 |
QEYVLTQQGFIYQGSAK/FLKNAGR | 2747.42 | 3+ | 677 | 164 | 8.31 × 10−11 | 4 |
QEYVLTQQGFIYQGSAK/TVEIPDPVEAGEEVKVR | 3808.93 | 6+ | 677 | 164 | 8.33 × 10−6 | 2 |
QEYVLTQQGFIYQGSAK/AVKGFR | 2619.36 | 4+ | 677 | 169 | 5.15 × 10−7 | 10 |
VVSGMVNCNDDQGVLLGR/AVKGFR e | 2535.28 | 4+ | 677 | 234 | 1.70 × 10−11 | 14 |
WKNHGCQR/AVKGFR | 1687.86 | 4+ | 677 | 270 | 1.80 × 10−5 | 3 |
VVTNYNSAHDQNSNLLIEYFR/AVKGFR e | 3156.59 | 4+ | 677 | 307 | 1.03 × 10−5 | 10 |
NEFGEIQGDK/AVKGFR | 1795.90 | 3+ | 677 | 324 | 1.37 × 10−8 | 5 |
VGQSMNMGSDFDVFAHITNNTAEEYVCR/AVKGFR e | 3794.74 | 4+ | 677 | 481 | 1.99 × 10−5 | 5 |
(A) Isopeptides with Unambiguous Modification Sites (Localization Probability >75%) | |||||||||||||
Position of K in TG2 a | m/z | z | Score b | Isopeptide Probability [%] | Deamidation Probability [%] | pLink2 Identification c | |||||||
Q2 | Q4 | Q9 | Q11 | Q2 | Q4 | Q9 | Q11 | E-value | No. MS2 Spectra | ||||
205 | 818.77 | 3+ | 176.20 | - | - | 0.7 | 99.3 | 12.8 | 87.2 | - | - | 1.49 × 10−9 | 15 |
380 | 904.13 | 3+ | 153.63 | - | - | 0.4 | 99.6 | - | 100 | - | - | 1.37 × 10−9 | 8 |
464 | 929.48 | 3+ | 158.40 | - | - | 0.7 | 99.3 | - | 100 | - | - | 3.30 × 10−6 | 9 |
590 | 1051.22 | 3+ | 126.76 | 8.7 | 91.3 | - | - | - | - | 0.1 | 99.9 | 1.30 × 10−11 | 3 |
1051.55 | 3+ | 72.29 | 1.1 | 7.8 | 90.8 | 0.3 | 89.1 | 10.9 | 0.4 | 99.6 | 1.95 × 10−11 | 5 | |
598 | 854.46 | 3+ | 134.18 | - | - | 0.3 | 99.7 | 0.3 | 99.7 | - | - | 2.11 × 10−11 | 14 |
600c | 1104.59 | 2+ | 165.83 | - | - | - | 100 | - | 100 | - | - | - | - |
649 | 1172.60 | 3+ | 64.65 | - | 0.1 | 2.1 | 97.8 | 10.5 | 89.4 | - | 0.1 | 2.00 × 10−4 | 7 |
672 | 980.85 | 3+ | 138.94 | - | - | 6.2 | 93.8 | 6.4 | 93.6 | - | - | 1.36 × 10−9 | 3 |
677 | 776.08 | 3+ | 84.29 | 8.1 | 89.4 | 0.5 | 2.1 | 0.2 | 2.1 | 0.9 | 96.8 | 2.85 × 10−5 | 3 |
(B) Isopeptides with Ambiguous Modification Sites | |||||||||||||
464 | 697.61 | 4+ | 61.64 | 0.9 | 0.8 | 91.7 | 6.5 | 49.9 | 50.1 | 6.8 | 93.2 | 1.46 × 10−3 | 6 |
468 | 1005.49 | 3+ | 43.99 | 0.9 | 4.6 | 50.6 | 43.9 | 10.0 | 84.3 | 1.0 | 4.6 | - | - |
1005.49 | 3+ | 40.73 | 0.2 | 2.2 | 19.3 | 78.3 | 41.8 | 55.2 | 0.8 | 2.2 | - | - | |
562 | 1013.85 | 3+ | 53.26 | - | 0.2 | 11.5 | 88.3 | 29.9 | 69.8 | 0.1 | 0.2 | 1.36 × 10−3 | 2 |
590 | 1051.22 | 3+ | 158.30 | - | - | 63.0 | 37.0 | - | 100 | - | - | - | - |
1051.22 | 3+ | 33.20 | 0.9 | 3.2 | 16.4 | 79.5 | 23.9 | 66.7 | 3.6 | 5.8 | 5.89 × 10−15 | 10 | |
600d | 694.03 | 3+ | 165.35 | - | - | 2.2 | 97.8 | 63.6 | 36.4 | - | - | - | - |
694.36 | 3+ | 44.62 | 10.2 | 28.8 | 23.8 | 37.3 | 76.3 | 62.1 | 52.0 | 9.6 | - | - | |
(C) Isopeptides Identified with pLink2 d | |||||||||||||
Crosslink | Deamidation | ||||||||||||
30 | 798.10 | 3+ | Q11 | Q4 | 1.20 × 10-4 | 3 | |||||||
797.77 | 3+ | Q4 | - | 7.62 × 10-5 | 3 | ||||||||
205 | 818.77 | 3+ | Q9 | Q4 | 7.67 × 10-3 | 6 | |||||||
429 | 832.77 | 3+ | Q11 | Q4 | 1.95 × 10-4 | 4 |
(A) Isopeptides with Unambiguous Modification Sites (Localization Probability >75%) | |||||||||||||||||
Position of K in TG2 a | m/z | z | Score b | Isopeptide Probability [%] | Deamidation Probability [%] | pLink2 Identification c | |||||||||||
Q2 | Q4 | Q8 | Q10 | Q11 | Q14 | Q2 | Q4 | Q8 | Q10 | Q11 | Q14 | E-Value | No. MS2 Spectra | ||||
205 | 798.76 | 3+ | 142.10 | 0.6 | 1.0 | 0.4 | 87.4 | 10.6 | - | 99.4 | 99.0 | - | 0.6 | - | - | - | - |
798.43 | 3+ | 139.74 | - | - | - | 99.6 | 0.4 | - | 0.5 | 99.5 | - | - | - | - | 2.35 × 10−6 | 5 | |
380 | 884.13 | 3+ | 125.61 | 90.7 | 8.1 | 1.2 | - | - | - | 8.3 | 91.7 | 0.2 | 98.2 | 1.6 | - | 9.80 × 10−7 | 2 |
884.13 | 3+ | 128.46 | - | - | 0.1 | 99.1 | 0.8 | - | 0.4 | 99.6 | - | - | - | - | 8.31 × 10−7 | 3 | |
464 | 682.12 | 4+ | 149.69 | - | - | - | 99.8 | 0.2 | - | 1.4 | 98.6 | - | - | - | - | 1.06 × 10−6 | 7 |
682.36 | 4+ | 131.69 | 89.6 | 10.4 | - | - | - | - | 10.4 | 88.3 | 1.3 | 98.7 | 1.3 | - | - | - | |
590 | 1031.22 | 3+ | 157.56 | 93.6 | 6.4 | - | - | - | - | 6.4 | 93.6 | - | 98.3 | 1.7 | - | 6.60 × 10−11 | 8 |
1031.22 | 3+ | 126.32 | 10.5 | 89.5 | - | - | - | - | 89.5 | 10.5 | 0.1 | 96.9 | 3.1 | - | 1.61 × 10−8 | 8 | |
1031.54 | 3+ | 71.88 | - | - | - | - | 0.1 | 99.9 | 100 | 99.9 | 0.5 | 85.3 | 14.2 | - | 3.53 × 10−9 | 4 | |
598 | 834.13 | 3+ | 126.32 | - | 0.9 | 14.7 | 83.8 | 0.5 | - | 0.6 | 98.5 | - | 0.9 | - | - | 3.65 × 10−3 | 2 |
834.79 | 3+ | 75.46 | 98.9 | 1.1 | - | - | - | - | 1.1 | 97.4 | 2.5 | 87.8 | 11.6 | 99.6 | - | - | |
600 | 1010.05 | 2+ | 232.88 | - | - | - | 100 | - | - | 0.1 | 99.9 | - | - | - | - | - | - |
674.03 | 3+ | 139.19 | 99.2 | 0.8 | - | - | - | - | 0.8 | 99.2 | - | 93.3 | 6.7 | - | - | - | |
649 | 1151.95 | 3+ | 52.82 | - | - | - | 0.2 | 1.9 | 97.9 | - | - | - | - | - | - | - | - |
1152.60 | 3+ | 51.55 | 0.5 | 2.6 | 3.0 | 2.8 | 77.2 | 13.9 | 86.3 | 13.1 | 1.9 | 94.6 | 3.8 | 0.4 | - | - | |
672 | 960.85 | 3+ | 67.02 | 96.7 | 3.1 | 0.2 | - | - | - | 3.1 | 95.7 | 1.5 | 95.9 | 3.8 | - | - | - |
677 | 756.08 | 3+ | 120.60 | 93.0 | 7.0 | - | - | - | - | 7.0 | 92.9 | 0.1 | 96.3 | 3.7 | - | 2.12 × 10−5 | 5 |
756.08 | 3+ | 115.12 | 98.5 | 1.5 | - | - | - | - | 1.4 | 98.5 | 10.9 | 89.0 | 0.2 | - | 1.33 × 10−6 | 4 | |
755.75 | 3+ | 60.18 | - | - | 1.4 | 76.6 | 21.9 | 0.1 | 0.6 | 99.0 | 0.3 | 0.1 | - | - | 4.29 × 10−5 | 3 | |
(B) Isopeptides with Ambiguous Modification Sites | |||||||||||||||||
649 | 1152.60 | 3+ | 78.53 | 58.2 | 14.9 | 26.0 | 0.8 | 0.1 | - | 35.0 | 62.1 | 3.0 | 77.2 | 22.7 | 0.1 | 9.03 × 10−4 | 3 |
1152.93 | 3+ | 60.40 | 3.3 | 30.0 | 5.0 | 60.3 | 0.4 | 0.9 | 96.1 | 66.5 | 2.2 | 35.9 | 8.7 | 90.5 | - | - | |
(C) Isopeptides Identified with pLink2 d | |||||||||||||||||
Crosslink | Deamidation | ||||||||||||||||
30 | 778.10 | 3+ | Q2 | Q4; Q10 | 8.12 × 10−5 | 3 | |||||||||||
778.10 | 3+ | Q4 | Q2; Q10 | 3.33 × 10−5 | 2 | ||||||||||||
380 | 884.46 | 3+ | Q2 | Q4; Q10; Q14 | 1.59 × 10−3 | 4 | |||||||||||
429 | 812.77 | 3+ | Q10 | Q2; Q4 | 4.79 × 10−5 | 6 | |||||||||||
464 | 909.15 | 3+ | Q11 | Q4 | 5.29 × 10−6 | 2 | |||||||||||
682.12 | 4+ | Q4 | Q10 | 4.42 × 10−7 | 5 | ||||||||||||
590 | 1031.22 | 3+ | Q8 | Q4; Q10 | 2.46 × 10−6 | 3 | |||||||||||
649 | 1152.60 | 3+ | Q11 | Q4; Q10 | 1.07 × 10−5 | 4 | |||||||||||
1152.60 | 3+ | Q8 | Q4; Q10 | 1.72 × 10−6 | 2 | ||||||||||||
1152.93 | 3+ | Q14 | Q2; Q10; Q14 | 2.12 × 10−6 | 2 | ||||||||||||
672 | 960.52 | 3+ | Q10 | Q4 | 2.81 × 10−4 | 2 | |||||||||||
960.52 | 3+ | Q4 | Q10 | 1.01 × 10−3 | 2 |
(A) Isopeptides with Unambiguous Modification Sites (Localization Probability >75%) | |||||||||||||||||||||||
Position of K in TG2 a | m/z | z | Score b | Isopeptide Probability [%] | Deamidation Probability [%] | pLink2 Identification c | |||||||||||||||||
Q2 | Q4 | Q5 | Q7 | Q8 | Q12 | Q13 | Q14 | Q15 | Q2 | Q4 | Q5 | Q7 | Q8 | Q12 | Q13 | Q14 | Q15 | E-value | No. MS2 Spectra | ||||
205 | 937.47 | 3+ | 184.99 | - | 80.3 | 11.9 | 7.8 | 0.1 | - | - | - | - | - | 7.4 | 3.7 | 88.6 | 0.2 | 80.2 | 28.7 | 91.0 | - | - | - |
590 | 1169.93 | 3+ | 133.23 | 4.9 | 90.1 | 4.9 | - | - | - | - | - | - | - | - | - | - | - | 99.3 | 16.4 | 82.1 | 2.2 | - | - |
600 | 812.74 | 3+ | 150.76 | - | 83.8 | 12.4 | 3.2 | 0.5 | - | - | - | - | - | 2.2 | 4.1 | 81.6 | 12.1 | 86.1 | 33.7 | 80.2 | - | ||
649 | 1291.31 | 3+ | 83.37 | 0.6 | 77.8 | 19.4 | 1.7 | 0.5 | - | - | - | - | 0.1 | 1.1 | 1.9 | 77.6 | 19.5 | 90.9 | 75.8 | 24.1 | 0.9 | - | - |
677 | 894.46 | 3+ | 140.22 | 0.3 | 87.4 | 11.9 | 0.3 | - | - | - | - | - | - | - | - | - | - | 98.4 | 16.8 | 84.6 | 0.2 | ||
(B) Isopeptides with Ambiguous Modification Sites | |||||||||||||||||||||||
205 | 937.46 | 3+ | 169.37 | - | - | - | - | - | 76.3 | 22.8 | 0.9 | - | - | 92.8 | 7.6 | 24.3 | 75.2 | 1.0 | 2.2 | 96.8 | - | 1.49 × 10−5 | 7 |
937.46 | 3+ | 153.48 | 0.1 | 31.0 | 4.6 | 63.8 | 0.5 | - | - | - | - | - | 31.1 | 34.5 | 34.0 | 0.5 | 73.6 | 31.2 | 94.6 | 0.5 | 9.00 × 10−5 | 5 | |
380 | 1022.85 | 3+ | 153.23 | - | 21.0 | 2.6 | 76.2 | 0.2 | - | - | - | - | - | 32.4 | 48.5 | 19.0 | 0.1 | 82.7 | 29.5 | 85.8 | 2.1 | 1.82 × 10−4 | 6 |
464 | 1048.20 | 3+ | 104.94 | - | 23.9 | 18.4 | 27.4 | 27.2 | 2.5 | 0.3 | 0.3 | - | - | 2.6 | 20.3 | 29.0 | 28.9 | 94.4 | 53.1 | 48.2 | 0.1 | - | - |
786.64 | 4+ | 103.88 | 0.4 | 72.8 | 13.1 | 12.6 | 1.2 | - | - | - | - | 7.6 | 8.9 | 37.4 | 39.1 | 6.9 | 94.5 | 44.7 | 57.3 | 3.5 | - | - | |
786.64 | 4+ | 71.55 | - | 0.1 | 1.2 | 76.3 | 19.6 | 0.8 | 0.5 | 0.5 | 0.1 | 30.3 | 49.6 | 20.5 | 21.5 | 78.9 | 37.8 | 75.1 | 70.9 | 15.3 | 4.86 × 10−4 | 4 | |
590 | 1169.60 | 3+ | 147.56 | - | 12.4 | 12.4 | 67.5 | 7.7 | - | - | - | - | 0.2 | 53.2 | 21.9 | 21.1 | 3.6 | 60.7 | 84.4 | 53.8 | 1.2 | 4.70 × 10−9 | 2 |
598 | 973.17 | 3+ | 148.70 | - | 9.2 | 1.5 | 44.6 | 44.6 | - | - | - | - | - | 38.4 | 51.4 | 5.1 | 5.1 | 96.4 | 98.7 | 4.8 | - | - | - |
973.17 | 3+ | 140.77 | - | 30.6 | 6.2 | 60.6 | 2.5 | 0.1 | 0.1 | - | - | - | 32.9 | 32.4 | 33.4 | 1.5 | 87.0 | 95.0 | 17.8 | - | 1.15 × 10−3 | 4 | |
972.84 | 3+ | 125.50 | - | 45.6 | 45.6 | 8.6 | 0.2 | - | - | - | - | - | - | - | - | - | 48.7 | 76.9 | 70.7 | 3.6 | 8.06 × 10−4 | 2 | |
600 | 812.74 | 3+ | 124.50 | - | 14.4 | 81.1 | 3.3 | 1.2 | - | - | - | - | 0.1 | 78.7 | 11.2 | 7.9 | 2.1 | 89.7 | 55.1 | 55.1 | 0.1 | - | - |
649 | 1291.64 | 3+ | 78.33 | - | 0.4 | 3.4 | 21.1 | 70.9 | 0.4 | 0.2 | - | - | 0.6 | 49.0 | 54.1 | 72.3 | 23.5 | 82.3 | 50.7 | 52.7 | 14.8 | 2.85 × 10−5 | 2 |
1291.64 | 3+ | 78.33 | - | 0.4 | 3.4 | 21.1 | 70.9 | 0.4 | 0.2 | - | - | 0.6 | 49.0 | 54.1 | 72.3 | 23.5 | 82.3 | 50.7 | 52.7 | 14.8 | 1.39 × 10−5 | 2 | |
1291.64 | 3+ | 60.54 | 1.4 | 68.1 | 27.5 | 2.3 | 0.5 | 0.1 | - | - | - | - | 0.1 | 0.2 | 0.6 | 5.6 | 71.3 | 36.5 | 53.9 | 31.6 | 2.63 × 10−6 | 3 | |
677 | 894.79 | 3+ | 178.14 | - | 6.4 | 1.0 | 89.3 | 3.4 | - | - | - | - | - | 36.5 | 56.6 | 6.4 | 0.4 | 99.8 | 99.8 | 0.4 | - | - | - |
894.46 | 3+ | 120.59 | - | 5.1 | 70.1 | 23.3 | 1.2 | 0.1 | 0.1 | 0.1 | - | - | 0.1 | 0.1 | - | - | 98.8 | 49.4 | 49.4 | 2.1 | - | - | |
894.45 | 3+ | 51.70 | - | 0.1 | 0.1 | 0.4 | 0.4 | 24.7 | 24.7 | 24.7 | 24.8 | 0.3 | 47.1 | 52.9 | 49.0 | 54.9 | 74.0 | 74.0 | 74.0 | 73.8 | - | - | |
(C) Isopeptides identified with pLink2 d | |||||||||||||||||||||||
Crosslink | Deamidation | ||||||||||||||||||||||
30 | 916.48 | 3+ | Q4 | Q12; Q14 | 4.14 × 10−4 | 3 | |||||||||||||||||
380 | 1022.52 | 3+ | Q5 | Q12; Q13 | 6.95 × 10−5 | 3 | |||||||||||||||||
429 | 951.15 | 3+ | Q12 | Q4; Q14 | 6.77 × 10−5 | 2 | |||||||||||||||||
464 | 786.15 | 4+ | Q12 | Q4; Q13 | 3.16 × 10−4 | 6 | |||||||||||||||||
786.64 | 4+ | Q12 | Q2; Q10; Q14 | 3.39 × 10−4 | 3 | ||||||||||||||||||
590 | 1169.60 | 3+ | Q12 | Q4; Q7; Q13 | 1.21 × 10−6 | 2 | |||||||||||||||||
1169.60 | 3+ | Q7 | Q12; Q14 | 5.18 × 10−9 | 2 |
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Share and Cite
Lexhaller, B.; Ludwig, C.; Scherf, K.A. Comprehensive Detection of Isopeptides between Human Tissue Transglutaminase and Gluten Peptides. Nutrients 2019, 11, 2263. https://doi.org/10.3390/nu11102263
Lexhaller B, Ludwig C, Scherf KA. Comprehensive Detection of Isopeptides between Human Tissue Transglutaminase and Gluten Peptides. Nutrients. 2019; 11(10):2263. https://doi.org/10.3390/nu11102263
Chicago/Turabian StyleLexhaller, Barbara, Christina Ludwig, and Katharina A. Scherf. 2019. "Comprehensive Detection of Isopeptides between Human Tissue Transglutaminase and Gluten Peptides" Nutrients 11, no. 10: 2263. https://doi.org/10.3390/nu11102263