The critical amino acids of a nephritogenic epitope on human Goodpasture autoantigen for binding to HLA-DRB1*1501
| Autor: | Jian-nan Li, Xiao-yu Jia, Ming-Hui Zhao, Zhao Cui, Fang-jin Chen, Qiu-hua Gu |
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| Rok vydání: | 2017 |
| Předmět: |
Collagen Type IV
0301 basic medicine Anti-Glomerular Basement Membrane Disease Immunology Lysine Epitopes T-Lymphocyte Autoimmunity Peptide Autoantigens Protein Structure Secondary Epitope 03 medical and health sciences medicine Humans Molecular Biology B cell Cell Line Transformed Alanine chemistry.chemical_classification B-Lymphocytes Chemistry Tryptophan Amino acid 030104 developmental biology medicine.anatomical_structure Amino Acid Substitution Biochemistry Binding Sites Antibody HLA-DRB1 Chains Cysteine |
| Zdroj: | Molecular Immunology. 88:1-9 |
| ISSN: | 0161-5890 |
| Popis: | Background Anti-GBM disease is caused by autoimmunity to Goodpasture antigen on α3(IV)NC1 and had strong associations with HLA-DRB1*1501. Previous studies identified α3 127-148 (P14: TDIPPCPHGWISLWKGFSFIMF) as a T cell epitope. The present study was aimed to investigate the binding capacity of P14 to HLA-DRB1*1501 and the critical amino acids for this binding. Methods A line of EBV-transformed human B cells homozygous for HLA-DRB1*1501 was used to detect the binding capacity of peptides to HLA-DRB1*1501 using flow cytometry analysis. P14 was sequentially truncated into 8 peptides with 15 amino acids to identify the core binding motif. A set of alanine substituted peptides of P14-2 was then synthesized to identify its critical residues for binding to HLA-DRB1*1501. The structure of HLA-DR2b-Peptide-TCR complex was constructed by modeling to analyze the interaction of each amino acids of P14-2 with the HLA-DR2b molecule. Results P14 could bind to HLA-DRB1*1501 expressed on B cell surface. The N-terminus of P14 was the core binding motif and the truncated peptide P14-2 (DIPPCPHGWISLWKG) 128-142 had the strongest binding capacity. After sequential amino acid substitution, we found the binding capacity of P14-2 was completely lost by the substitution of cysteine (C) 132 and significantly decreased by the substitution of tryptophan (W) 136 , lysine (K) 141 , or glycine (G) 142 , but still at a high level. The modeling showed that (C) 132 had a strong interaction with pocket 4 on the β chain of DR2b. Thus, C 132 , W 136 , K 141 , and G 142 were defined as the critical amino acid residues for the binding capacity of P14 to HLA-DRB1*1501. Conclusion We identified α3 128-142 (DIPPCPHGWISLWKG) as the core binding motif of P14 to HLA-DRB1*1501 molecule. And the critical amino acid residues for this binding were further defined as C 132 , W 136 , K 141 , and G 142 . |
| Databáze: | OpenAIRE |
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