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Identification of novel interferon responsive protein partners of human leukocyte antigen A (HLA-A) using cross-linking mass spectrometry (CLMS) approach
A. Singh, M. Padariya, J. Faktor, S. Kote, S. Mikac, A. Dziadosz, TW. Lam, J. Brydon, MA. Wear, KL. Ball, T. Hupp, A. Sznarkowska, B. Vojtesek, U. Kalathiya
Language English Country England, Great Britain
Document type Journal Article, Research Support, Non-U.S. Gov't
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- MeSH
- HLA Antigens MeSH
- HLA-A Antigens MeSH
- Mass Spectrometry methods MeSH
- Interferons * MeSH
- Humans MeSH
- Proteins * chemistry MeSH
- Cross-Linking Reagents chemistry MeSH
- Ubiquitin Thiolesterase MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The interferon signalling system elicits a robust cytokine response against a wide range of environmental pathogenic and internal pathological signals, leading to induction of a subset of interferon-induced proteins. We applied DSS (disuccinimidyl suberate) mediated cross-linking mass spectrometry (CLMS) to capture novel protein-protein interactions within the realm of interferon induced proteins. In addition to the expected interferon-induced proteins, we identified novel inter- and intra-molecular cross-linked adducts for the canonical interferon induced proteins, such as MX1, USP18, OAS3, and STAT1. We focused on orthogonal validation of a cohort of novel interferon-induced protein networks formed by the HLA-A protein (H2BFS-HLA-A-HMGA1) using co-immunoprecipitation assay, and further investigated them by molecular dynamics simulation. Conformational dynamics of the simulated protein complexes revealed several interaction sites that mirrored the interactions identified in the CLMS findings. Together, we showcase a proof-of-principle CLMS study to identify novel interferon-induced signaling complexes and anticipate broader use of CLMS to identify novel protein interaction dynamics within the tumour microenvironment.
Institute of Genetics and Cancer University of Edinburgh Edinburgh EH4 2XR Scotland UK
RECAMO Masaryk Memorial Cancer Institute Zlutykopec 7 65653 Brno Czech Republic
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