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Title: Fluorogenic Probes for Monitoring Peptide Binding to Class II MHC Proteins in Living Cells

Abstract

A crucial step in the immune response is the binding of antigenic peptides to major histocompatibility complex (MHC) proteins. Class II MHC proteins present their bound peptides to CD4+ T cells, thereby helping to activate both the humoral and the cellular arms of the adaptive immune response. Peptide loading onto class II MHC proteins is regulated temporally, spatially and developmentally in antigen-presenting cells1. To help visualize these processes, we have developed a series of novel fluorogenic probes that incorporate the environment-sensitive amino acid analogs 6-N,N-dimethylamino-2-3-naphthalimidoalanine and 4-N,N-dimethylaminophthalimidoalanine. Upon binding to class II MHC proteins these fluorophores show large changes in emission spectra, quantum yield and fluorescence lifetime. Peptides incorporating these fluorophores bind specifically to class II MHC proteins on antigen-presenting cells and can be used to follow peptide binding in vivo. Using these probes we have tracked a developmentally regulated cell-surface peptide-binding activity in primary human monocyte-derived dendritic cells.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
930650
Report Number(s):
BNL-81094-2008-JA
TRN: US200901%%162
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Nature Chemical Biology; Journal Volume: 3; Journal Issue: 4
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; AMINO ACIDS; EMISSION SPECTRA; FLUORESCENCE; HISTOCOMPATIBILITY COMPLEX; IN VIVO; LIFETIME; MONITORING; PEPTIDES; PROBES; PROTEINS; national synchrotron light source

Citation Formats

Venkatraman,P., Nguyen, T., Sainlos, M., Bilsel, o., Chitta, S., Imperiali, B., and Stern, L. Fluorogenic Probes for Monitoring Peptide Binding to Class II MHC Proteins in Living Cells. United States: N. p., 2007. Web. doi:10.1038/nchembio868.
Venkatraman,P., Nguyen, T., Sainlos, M., Bilsel, o., Chitta, S., Imperiali, B., & Stern, L. Fluorogenic Probes for Monitoring Peptide Binding to Class II MHC Proteins in Living Cells. United States. doi:10.1038/nchembio868.
Venkatraman,P., Nguyen, T., Sainlos, M., Bilsel, o., Chitta, S., Imperiali, B., and Stern, L. Mon . "Fluorogenic Probes for Monitoring Peptide Binding to Class II MHC Proteins in Living Cells". United States. doi:10.1038/nchembio868.
@article{osti_930650,
title = {Fluorogenic Probes for Monitoring Peptide Binding to Class II MHC Proteins in Living Cells},
author = {Venkatraman,P. and Nguyen, T. and Sainlos, M. and Bilsel, o. and Chitta, S. and Imperiali, B. and Stern, L.},
abstractNote = {A crucial step in the immune response is the binding of antigenic peptides to major histocompatibility complex (MHC) proteins. Class II MHC proteins present their bound peptides to CD4+ T cells, thereby helping to activate both the humoral and the cellular arms of the adaptive immune response. Peptide loading onto class II MHC proteins is regulated temporally, spatially and developmentally in antigen-presenting cells1. To help visualize these processes, we have developed a series of novel fluorogenic probes that incorporate the environment-sensitive amino acid analogs 6-N,N-dimethylamino-2-3-naphthalimidoalanine and 4-N,N-dimethylaminophthalimidoalanine. Upon binding to class II MHC proteins these fluorophores show large changes in emission spectra, quantum yield and fluorescence lifetime. Peptides incorporating these fluorophores bind specifically to class II MHC proteins on antigen-presenting cells and can be used to follow peptide binding in vivo. Using these probes we have tracked a developmentally regulated cell-surface peptide-binding activity in primary human monocyte-derived dendritic cells.},
doi = {10.1038/nchembio868},
journal = {Nature Chemical Biology},
number = 4,
volume = 3,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}