Application of the immunoregulatory receptor LILRB1 as a novel crystallisation chaperone for class I peptide-MHC complexes
| Autor: | Benjamin E. Willcox, Daniel H. Stones, Fiyaz Mohammed |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences biology chemical and pharmacologic phenomena Peptide Computational biology Protein engineering Major histocompatibility complex LILRB1 03 medical and health sciences 0302 clinical medicine chemistry Chaperone (protein) Immunology biology.protein Peptide-MHC Receptor Antigenic peptide 030304 developmental biology 030215 immunology |
| DOI: | 10.1101/213207 |
| Popis: | X-ray crystallographic studies of class I peptide-MHC molecules (pMHC) continue to provide important insights into immune recognition, however their success depends on generation of diffraction-quality crystals, which remains a significant challenge. While protein engineering techniques such as surface-entropy reduction and lysine methylation have proven utility in facilitating and/or improving protein crystallisation, they risk affecting the conformation and biochemistry of the class I MHC antigen binding groove. An attractive alternative is the use of noncovalent crystallisation chaperones, however these have not been developed for pMHC. Here we describe a method for promoting class I pMHC crystallisation, by exploiting its natural ligand interaction with the immunoregulatory receptor LILRB1 as a novel crystallisation chaperone. First, focussing on a model HIV-1-derived HLA-A2-restricted peptide, we determined a 2.4Å HLA-A2/LILRB1 structure, which validated that co-crystallisation with LILRB1 does not alter conformation of the antigenic peptide. We then demonstrated that addition of LILRB1 enhanced the crystallisation of multiple pMHC complexes, and identified a generic condition for initial co-crystallisation. LILRB1 chaperone-based pMHC crystallisation enabled structure determination for class I pMHC previously intransigent to crystallisation, including both conventional and post-translationally-modified peptides, of diverse lengths. LILRB1 chaperone-mediated crystallisation should expedite molecular insights into the immunobiology of diverse immune-related diseases and immunotherapeutic strategies, particularly involving class I pMHC complexes that are challenging to crystallise. Moreover, since the LILRB1 recognition interface involves predominantly non-polymorphic regions of the MHC molecule, the approach we outline could prove applicable to a diverse range of class I pMHC. |
| Databáze: | OpenAIRE |
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