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Expanding the detectable HLA peptide repertoire using electron-transfer/higher-energy collision dissociation (EThcD).


ABSTRACT: The identification of peptides presented by human leukocyte antigen (HLA) class I is tremendously important for the understanding of antigen presentation mechanisms under healthy or diseased conditions. Currently, mass spectrometry-based methods represent the best methodology for the identification of HLA class I-associated peptides. However, the HLA class I peptide repertoire remains largely unexplored because the variable nature of endogenous peptides represents difficulties in conventional peptide fragmentation technology. Here, we substantially enhanced (about threefold) the identification success rate of peptides presented by HLA class I using combined electron-transfer/higher-energy collision dissociation (EThcD), reporting over 12,000 high-confident (false discovery rate <1%) peptides from a single human B-cell line. The direct importance of such an unprecedented large dataset is highlighted by the discovery of unique features in antigen presentation. The observation that a substantial part of proteins is sampled across different HLA alleles, and the common occurrence of HLA class I nested sets, suggest that the constraints of HLA class I to comprehensively present the health states of cells are not as tight as previously thought. Our dataset contains a substantial set of peptides bearing a variety of posttranslational modifications presented with marked allele-specific differences. We propose that EThcD should become the method of choice in analyzing HLA class I-presented peptides.

SUBMITTER: Mommen GP 

PROVIDER: S-EPMC3970485 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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Expanding the detectable HLA peptide repertoire using electron-transfer/higher-energy collision dissociation (EThcD).

Mommen Geert P M GP   Frese Christian K CK   Meiring Hugo D HD   van Gaans-van den Brink Jacqueline J   de Jong Ad P J M AP   van Els Cécile A C M CA   Heck Albert J R AJ  

Proceedings of the National Academy of Sciences of the United States of America 20140310 12


The identification of peptides presented by human leukocyte antigen (HLA) class I is tremendously important for the understanding of antigen presentation mechanisms under healthy or diseased conditions. Currently, mass spectrometry-based methods represent the best methodology for the identification of HLA class I-associated peptides. However, the HLA class I peptide repertoire remains largely unexplored because the variable nature of endogenous peptides represents difficulties in conventional pe  ...[more]

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