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

Data from: TAPBPR alters MHC class I peptide presentation by functioning as a peptide exchange catalyst
Data from: TAPBPR alters MHC class I peptide presentation by functioning as a peptide exchange catalyst
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,
DRYAD
Hermann, Clemens
978cedee-4017-43e9-86a7-4665636ce553
Van Hateren, Andy
e345fa3c-d89c-4b91-947e-c1d818cc7f71
Trautwein, Nico
2387bf8d-9384-4edb-93e7-3cefe5021ae7
Neerincx, Andreas
e487d7b5-1d13-4847-83b2-67fc2754c238
Duriez, Patrick J.
4cf499bc-007a-43b3-b180-d6e5dc3d151b
Stevanović, Stefan
e09af639-b73f-459b-b111-55a6c722b03d
Trowsdale, John
5bbacc4a-45f4-4376-8357-a7c79fc4fe12
Deane, Janet E.
dd93a448-fe81-4c8b-884e-a396ada0cebd
Elliott, Tim
16670fa8-c2f9-477a-91df-7c9e5b453e0e
Boyle, Louise H.
e2afc84e-3524-417d-a54b-4f7f7d753866
Hermann, Clemens
978cedee-4017-43e9-86a7-4665636ce553
Van Hateren, Andy
e345fa3c-d89c-4b91-947e-c1d818cc7f71
Trautwein, Nico
2387bf8d-9384-4edb-93e7-3cefe5021ae7
Neerincx, Andreas
e487d7b5-1d13-4847-83b2-67fc2754c238
Duriez, Patrick J.
4cf499bc-007a-43b3-b180-d6e5dc3d151b
Stevanović, Stefan
e09af639-b73f-459b-b111-55a6c722b03d
Trowsdale, John
5bbacc4a-45f4-4376-8357-a7c79fc4fe12
Deane, Janet E.
dd93a448-fe81-4c8b-884e-a396ada0cebd
Elliott, Tim
16670fa8-c2f9-477a-91df-7c9e5b453e0e
Boyle, Louise H.
e2afc84e-3524-417d-a54b-4f7f7d753866

Hermann, Clemens, Trautwein, Nico, Neerincx, Andreas, Stevanović, Stefan, Trowsdale, John, Deane, Janet E. and Boyle, Louise H. (2015) Data from: TAPBPR alters MHC class I peptide presentation by functioning as a peptide exchange catalyst. DRYAD doi:10.5061/dryad.487j9 [Dataset]

Record type: 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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Published date: 1 January 2015

Identifiers

Local EPrints ID: 448551
URI: http://eprints.soton.ac.uk/id/eprint/448551
PURE UUID: 3df39df5-84b7-4354-8b60-24736997f6b4
ORCID for Andy Van Hateren: ORCID iD orcid.org/0000-0002-3915-0239
ORCID for Tim Elliott: ORCID iD orcid.org/0000-0003-1097-0222

Catalogue record

Date deposited: 26 Apr 2021 18:36
Last modified: 27 Apr 2021 01:41

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Contributors

Creator: Clemens Hermann
Contributor: Andy Van Hateren ORCID iD
Creator: Nico Trautwein
Creator: Andreas Neerincx
Contributor: Patrick J. Duriez
Creator: Stefan Stevanović
Creator: John Trowsdale
Creator: Janet E. Deane
Contributor: Tim Elliott ORCID iD
Creator: Louise H. Boyle

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