Endolysosomal Degradation of Allergenic Ole e 1-Like Proteins: Analysis of Proteolytic Cleavage Sites Revealing T Cell Epitope-Containing Peptides
Abstract
:1. Introduction
2. Results
2.1. Ole e 1-Like Allergens Showed Different Susceptibility to Endolysosomal Degradation
2.2. Structural Modeling and Molecular Dynamics Simulation
2.3. Endolysosomal Cleavage Sites
2.4. Substrate Binding Studies
2.5. Cathepsin S Inhibition
3. Discussion
4. Materials and Methods
4.1. Recombinant Production of Ole e 1-Like Allergens
4.2. Amino Acid Analysis
4.3. Endolysosomal Degradation Assay
4.4. Cathepsin S Inhibition Assays
4.5. Gel Electrophoresis
4.6. T Cell Epitope Prediction
4.7. Mass Spectrometry
4.8. Homology Modeling and Molecular Dynamics
4.9. Structure Preparation and Docking
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
ACE | Acetyl |
DC | Dendritic cell |
HCl | Hydrochloric acid |
MHC | Major histocompatibility complex |
NME | N-methyl |
SDS-PAGE | Sodium dodecyl sulfate polyacrylamide gel electrophoresis |
Appendix A
Amino Acid | P4 | P3 | P2 | P1 | P1′ | P2′ | P3′ | P4′ |
---|---|---|---|---|---|---|---|---|
Ala | 63 | 70 | 22 | 87 | 107 | 98 | 82 | 53 |
Arg | 6 | 7 | 1 | 13 | 6 | 12 | 8 | 8 |
Asn | 30 | 34 | 4 | 41 | 28 | 30 | 39 | 30 |
Asp | 37 | 19 | 0 | 28 | 33 | 27 | 71 | 77 |
Cys | 9 | 20 | 14 | 14 | 15 | 19 | 6 | 14 |
Gln | 38 | 33 | 14 | 82 | 67 | 51 | 37 | 47 |
Glu | 59 | 53 | 2 | 81 | 68 | 38 | 42 | 80 |
Gly | 48 | 63 | 5 | 128 | 118 | 86 | 80 | 91 |
His | 20 | 17 | 3 | 17 | 26 | 29 | 55 | 40 |
Ile | 25 | 58 | 76 | 9 | 19 | 34 | 21 | 27 |
Leu | 40 | 71 | 244 | 44 | 24 | 31 | 23 | 22 |
Lys | 7 | 9 | 1 | 8 | 6 | 26 | 15 | 23 |
Met | 20 | 20 | 55 | 15 | 10 | 8 | 2 | 6 |
Phe | 30 | 42 | 69 | 10 | 14 | 16 | 22 | 12 |
Pro | 42 | 51 | 5 | 1 | 0 | 63 | 106 | 79 |
Ser | 51 | 57 | 11 | 85 | 128 | 72 | 51 | 51 |
Thr | 22 | 41 | 17 | 76 | 40 | 59 | 39 | 46 |
Trp | 10 | 7 | 11 | 6 | 4 | 4 | 8 | 4 |
Tyr | 14 | 14 | 28 | 5 | 13 | 6 | 16 | 10 |
Val | 48 | 47 | 172 | 6 | 24 | 41 | 27 | 30 |
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Allergen | Plant | Accession Number | Host Organism | Protein Modification |
---|---|---|---|---|
Ole e 1.0101 | Olea europaea | P19963.2 | P. pastoris | A99V, K106L, N111Q, deamidation, oxidation |
Olive | ||||
Fra e 1.0101 | Fraxinus excelsior | AAQ08947.1 | P. pastoris | minor deamidations, yeast-specific glycosylation at N111 |
Ash | ||||
Sal k 5.0101 | Salsola kali | ADK22842.1 | P. pastoris | K3N, G84D, I91V, deamidation, yeast-specific glycosylation at N43 |
Russian thistle | ||||
Che a 1.0101 | Chenopodium album | Q8LGR0.1 | P. pastoris | deamidation, methionine oxidation, yeast-specific glycosylation at N39 |
Lamb’s quarters | ||||
Phl p 11.0101 | Phleum pratense | Q8H6L7.1 | P. pastoris | N24Q |
Timothy grass | ||||
Pla l 1.0101 | Plantago lanceolata | P82242.2 | E. coli | None |
English plantain |
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Wildner, S.; Elsässer, B.; Stemeseder, T.; Briza, P.; Soh, W.T.; Villalba, M.; Lidholm, J.; Brandstetter, H.; Gadermaier, G. Endolysosomal Degradation of Allergenic Ole e 1-Like Proteins: Analysis of Proteolytic Cleavage Sites Revealing T Cell Epitope-Containing Peptides. Int. J. Mol. Sci. 2017, 18, 1780. https://doi.org/10.3390/ijms18081780
Wildner S, Elsässer B, Stemeseder T, Briza P, Soh WT, Villalba M, Lidholm J, Brandstetter H, Gadermaier G. Endolysosomal Degradation of Allergenic Ole e 1-Like Proteins: Analysis of Proteolytic Cleavage Sites Revealing T Cell Epitope-Containing Peptides. International Journal of Molecular Sciences. 2017; 18(8):1780. https://doi.org/10.3390/ijms18081780
Chicago/Turabian StyleWildner, Sabrina, Brigitta Elsässer, Teresa Stemeseder, Peter Briza, Wai Tuck Soh, Mayte Villalba, Jonas Lidholm, Hans Brandstetter, and Gabriele Gadermaier. 2017. "Endolysosomal Degradation of Allergenic Ole e 1-Like Proteins: Analysis of Proteolytic Cleavage Sites Revealing T Cell Epitope-Containing Peptides" International Journal of Molecular Sciences 18, no. 8: 1780. https://doi.org/10.3390/ijms18081780