Application of the immunoregulatory receptor LILRB1 as a crystallisation chaperone for human class I MHC complexes

Mohammed, Fiyaz, Stones, Daniel H ORCID: 0000-0002-8981-7943 and Willcox, Benjamin E. (2019) Application of the immunoregulatory receptor LILRB1 as a crystallisation chaperone for human class I MHC complexes. Journal of Immunological Methods, 464. pp. 47-56. doi:10.1016/j.jim.2018.10.011

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Abstract

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 2.4 Å 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 peptide-HLA-A2 complexes, and identified a generic condition for initial co-crystallisation. LILRB1 chaperone-based crystallisation enabled structure determination for HLA-A2 complexes previously intransigent to crystallisation, including both conventional and post-translationally-modified peptides, of diverse lengths. Since both the LILRB1 recognition interface on the HLA-A2 α3 domain molecule and HLA-A2 mediated crystal contacts are predominantly conserved across class I MHC molecules, the approach we outline could prove applicable to a diverse range of class I pMHC. 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.

Item Type:	Article
Article Type:	Article
Uncontrolled Keywords:	Immunobiology; Immune-related diseases
Subjects:	Q Science > Q Science (General) R Medicine > RB Pathology
Divisions:	Schools and Research Institutes > School of Education and Science
Research Priority Areas:	Place, Environment and Community
Depositing User:	Daniel Stones
Date Deposited:	01 Nov 2018 13:43
Last Modified:	01 Sep 2023 15:29
URI:	https://eprints.glos.ac.uk/id/eprint/6148

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