Data from: TAPBPR alters MHC class I peptide presentation by functioning as a peptide exchange catalyst

(2015) Data from: TAPBPR alters MHC class I peptide presentation by functioning as a peptide exchange catalyst. DRYAD doi:10.5061/dryad.487j9 [Dataset]

Abstract

Our understanding of the antigen presentation pathway has recently been enhanced with the identification that the tapasin-related protein TAPBPR is a second MHC I-specific chaperone. We sought to determine whether, like tapasin, TAPBPR can also influence MHC I peptide selection by functioning as a peptide exchange catalyst. We show that TAPBPR can catalyse the dissociation of peptides from peptide-MHC I complexes, enhance the loading of peptide-receptive MHC I molecules, and discriminate between peptides based on affinity in vitro. In cells, the depletion of TAPBPR increased the diversity of peptides presented on MHC I molecules, suggesting that TAPBPR is involved in restricting peptide presentation. Our results suggest TAPBPR binds to MHC I in a peptide-receptive state and, like tapasin, works to enhance peptide optimisation. It is now clear there are two MHC class I specific peptide editors, tapasin and TAPBPR, intimately involved in controlling peptide presentation to the immune system.,Figure 5C - Peptides eluted from MHC class I molecules in Hela+IFNy and Hela-TAPBPR KO+IFNyPeptide-MHC class I complexes were isolated by affinity chromatography using W6/32 from IFN-γ treated HeLa and HeLa-TAPBPR KO cells. Eluted peptides were analysed using LC-MS/MS. Peptides were assigned as HLA-A*68:02 and HLA-B*15:03 binders based on their peptide motifs using the online programme NetMHC.Hela IFNy and Hela IFNy TAPBPR KO.xlsxFigure 5D - peptides eluted from HeLa and HeLa-WT-TAPBPR cellsPeptide-MHC class I complexes were isolated by affinity chromatography using W6/32 from HeLa and HeLa overexpressing WT-TAPBPR. Eluted peptides were analysed using LC-MS/MS. Peptides were assigned as HLA-A*68:02 and HLA-B*15:03 binders based on their peptide motifs using the online programme NetMHC.HeLa and HeLa WT TAPBPR.xlsxFigure 5 - Figure supplement 1 - Peptides eluted from HeLa-S+IFNy and HeLa-S-shTAPBPR+IFNyPeptide-MHC class I complexes were isolated by affinity chromatography using W6/32 from IFN-γ treated HeLa-S cells and IFN-γ treated HeLa-S cells depleted of TAPBPR using shRNA. Eluted peptides were analysed using LC-MS/MS. Peptides were assigned as HLA-A*68:02 and HLA-B*15:03 binders based on their peptide motifs using the online programme NetMHC.HeLa IFNy and HeLa INFy shTAPBPR.xlsxFigure 6B - Peptide eluted from HLA-A2 from WT, TAPBPR depleted, and tapasin deficient KBM-7 cellsPeptide loaded HLA-A2 complexes were isolated using affinity chromatography with BB7.2 from IFN-γ treated WT, TAPBPR stably depleted (shTAPBPR) and tapasin deficient (TPN-) KBM-7 cells. Eluted peptide were analysed using LC-MS/MS.KBM7 elution from BB72.xlsxFigure 6E - Peptides eluted from HLA-B & -C from WT, TAPBPR depleted and tapasin deficient KBM-7 cellsHLA-B and -C complexes were isolated using affinity chromatography using B1.23.2 from IFN-γ treated WT, TAPBPR stably depleted (shTAPBPR) and tapasin deficient (TPN-) KBM-7 cells. Eluted peptide were analysed using LC-MS/MS.KBM7 elution from B1232.xlsx

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Contributors

Contributor: Andy Van Hateren

Contributor: Patrick J. Duriez

Contributor: Tim Elliott

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