Stable isotope tagging of epitopes: a highly selective strategy for the identification of major histocompatibility complex class I-associated peptides induced upon viral infection.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
AuthorsMeiring, Hugo D
Soethout, Ernst C
Poelen, Martien C M
Boog, Claire J
Heck, Albert J R
Jong, Ad P J M de
Els, Cécile A C M van
MetadataShow full item record
TitleStable isotope tagging of epitopes: a highly selective strategy for the identification of major histocompatibility complex class I-associated peptides induced upon viral infection.
PubliekssamenvattingIdentification of peptides presented in major histocompatibility complex (MHC) class I molecules after viral infection is of strategic importance for vaccine development. Until recently, mass spectrometric identification of virus-induced peptides was based on comparative analysis of peptide pools isolated from uninfected and virus-infected cells. Here we report on a powerful strategy aiming at the rapid, unambiguous identification of naturally processed MHC class I-associated peptides, which are induced by viral infection. The methodology, stable isotope tagging of epitopes (SITE), is based on metabolic labeling of endogenously synthesized proteins during infection. This is accomplished by culturing virus-infected cells with stable isotope-labeled amino acids that are expected to be anchor residues (i.e. residues of the peptide that have amino acid side chains that bind into pockets lining the peptide-binding groove of the MHC class I molecule) for the human leukocyte antigen allele of interest. Subsequently these cells are mixed with an equal number of non-infected cells, which are cultured in normal medium. Finally peptides are acid-eluted from immunoprecipitated MHC molecules and subjected to two-dimensional nanoscale LC-MS analysis. Virus-induced peptides are identified through computer-assisted detection of characteristic, binomially distributed ratios of labeled and unlabeled molecules. Using this approach we identified novel measles virus and respiratory syncytial virus epitopes as well as infection-induced self-peptides in several cell types, showing that SITE is a unique and versatile method for unequivocal identification of disease-related MHC class I epitopes.
- Targeted identification of infection-related HLA class I-presented epitopes by stable isotope tagging of epitopes (SITE).
- Authors: Meiring HD, Soethout EC, de Jong APJM, van Els CACM
- Issue date: 2007 May
- A microcapillary column switching HPLC-electrospray ionization MS system for the direct identification of peptides presented by major histocompatibility complex class I molecules.
- Authors: van der Heeft E, ten Hove GJ, Herberts CA, Meiring HD, van Els CA, de Jong AP
- Issue date: 1998 Sep 15
- Rapid and sensitive identification of major histocompatibility complex class I-associated tumor peptides by Nano-LC MALDI MS/MS.
- Authors: Hofmann S, Glückmann M, Kausche S, Schmidt A, Corvey C, Lichtenfels R, Huber C, Albrecht C, Karas M, Herr W
- Issue date: 2005 Dec
- The canine MHC class Ia allele DLA-88*508:01 presents diverse self- and canine distemper virus-origin peptides of varying length that have a conserved binding motif.
- Authors: Ross P, Nemec PS, Kapatos A, Miller KR, Holmes JC, Suter SE, Buntzman AS, Soderblom EJ, Collins EJ, Hess PR
- Issue date: 2018 Mar
- Endogenous presentation of a nascent antigenic epitope to CD8+ CTL is more efficient than exogenous presentation.
- Authors: Hahn YS, Hahn CS, Braciale TJ
- Issue date: 1996 Oct