Non-natural and photo-reactive amino acids as biochemical probes of immune function Journal Article

   


Authors: Gómez-Nuñez, M.; Haro, K. J.; Dao, T.; Chau, D.; Won, A.; Escobar-Alvarez, S.; Zakhaleva, V.; Korontsvit, T.; Gin, D. Y.; Scheinberg, D. A.
Article Title: Non-natural and photo-reactive amino acids as biochemical probes of immune function
Abstract: Wilms tumor protein (WT1) is a transcription factor selectively overexpressed in leukemias and cancers; clinical trials are underway that use altered WT1 peptide sequences as vaccines. Here we report a strategy to study peptide-MHC interactions by incorporating non-natural and photo-reactive amino acids into the sequence of WT1 peptides. Thirteen WT1 peptides sequences were synthesized with chemically modified amino acids (via fluorination and photo-reactive group additions) at MHC and T cell receptor binding positions. Certain new non-natural peptide analogs could stabilize MHC class I molecules better than the native sequences and were also able to elicit specific T-cell responses and sometimes cytotoxicity to leukemia cells. Two photo-reactive peptides, also modified with a biotin handle for pull-down studies, formed covalent interactions with MHC molecules on live cells and provided kinetic data showing the rapid clearance of the peptide-MHC complex. Despite "infinite affinity" provided by the covalent peptide bonding to the MHC, immunogenicity was not enhanced by these peptides because the peptide presentation on the surface was dominated by catabolism of the complex and only a small percentage of peptide molecules covalently bound to the MHC molecules. This study shows that nonnatural amino acids can be successfully incorporated into T cell epitopes to provide novel immunological, biochemical and kinetic information. © 2008 Gómez-Nuñez et al.
Keywords: controlled study; unclassified drug; human cell; molecular genetics; t lymphocyte; metabolism; cell line; protein binding; protein interaction; cytotoxicity; in vitro study; drug effect; peptide; radiation exposure; t lymphocyte receptor; immunology; lymphocyte activation; chemistry; donor; tissue donors; immune response; amino acid sequence; molecular sequence data; human cell culture; kinetics; gamma interferon; immunogenicity; leukemia cell; cytotoxic t lymphocyte; t-lymphocytes, cytotoxic; western blotting; peptides; amino acid; immunoassay; immunity; interferon-gamma; wt1 protein; amino acids; peptide vaccine; hla a0201 antigen; enzyme linked immunospot assay; hla a antigen; molecular probe; hla-a antigens; nephroblastoma; major histocompatibility antigen class 1; catabolism; wt1 proteins; cross linking reagent; cross-linking reagents; light; major histocompatibility antigen; histocompatibility antigens; hla-a0201 antigen; histocompatibility antigen; fluorination; molecular probes
Journal Title: PLoS ONE
Volume: 3
Issue: 12
ISSN: 1932-6203
Publisher: Public Library of Science  
Date Published: 2008-01-01
Start Page: e3938
Language: English
DOI: 10.1371/journal.pone.0003938
PUBMED: 19079589
PROVIDER: scopus
PMCID: PMC2592539
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-58049208310&partnerID=40&md5=88acba0ee2409d6ef59a425c432a4856
Notes: --- - "Cited By (since 1996): 3" - "Export Date: 17 November 2011" - "Source: Scopus"
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MSK Authors
  1. Marta Gomez
    11 Gomez
  2. Deming Chau
    12 Chau
  3. David Scheinberg
    500 Scheinberg
  4. Tao Dao
    82 Dao
  5. David Y Gin
    41 Gin
  6. Sindy N Escobar Alvarez
    8 Escobar-Alvarez
  7. Annie Won
    4 Won
  8. Viktoriya Zakhaleva
    29 Zakhaleva
  9. Tatyana Korontsvit
    53 Korontsvit
  10. Kurtis Haro
    5 Haro
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