Monoclonal Antibodies Specific for the Empty Conformation of HLA-DR1 Reveal Aspects of the Conformational Change Associated with Peptide Binding

Gregory J. Carven, Sriram Chitta, Ivan Hilgert, Mia M. Rushe, Rick F. Baggio, Michelle Palmer, Jaime E. Arenas, Jack L. Strominger, Vaclav Horejsi, Laura Santambrogio, Lawrence J. Stern

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Class II major histocompatibility complex (MHC) proteins bind peptides and present them at the cell surface for interaction with CD4+ T cells as part of the system by which the immune system surveys the body for signs of infection. Peptide binding is known to induce conformational changes in class II MHC proteins on the basis of a variety of hydrodynamic and spectroscopic approaches, but the changes have not been clearly localized within the overall class II MHC structure. To map the peptide-induced conformational change for HLA-DR1, a common human class II MHC variant, we generated a series of monoclonal antibodies recognizing the β subunit that are specific for the empty conformation. Each antibody reacted with the empty but not the peptide-loaded form, for both soluble recombinant protein and native protein expressed at the cell surface. Antibody binding epitopes were characterized using overlapping peptides and alanine scanning substitutions and were localized to two distinct regions of the protein. The pattern of key residues within the epitopes suggested that the two epitope regions undergo substantial conformational alteration during peptide binding. These results illuminate aspects of the structure of the empty forms and the nature of the peptide-induced conformational change.

Original languageEnglish (US)
Pages (from-to)16561-16570
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number16
DOIs
StatePublished - Apr 16 2004

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HLA-DR1 Antigen
Conformations
Monoclonal Antibodies
Peptides
Major Histocompatibility Complex
Epitopes
Proteins
T-cells
Antibodies
Immune system
Hydrodynamics
Recombinant Proteins
Cell Communication
Alanine
Immune System
Substitution reactions
Scanning
T-Lymphocytes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Monoclonal Antibodies Specific for the Empty Conformation of HLA-DR1 Reveal Aspects of the Conformational Change Associated with Peptide Binding. / Carven, Gregory J.; Chitta, Sriram; Hilgert, Ivan; Rushe, Mia M.; Baggio, Rick F.; Palmer, Michelle; Arenas, Jaime E.; Strominger, Jack L.; Horejsi, Vaclav; Santambrogio, Laura; Stern, Lawrence J.

In: Journal of Biological Chemistry, Vol. 279, No. 16, 16.04.2004, p. 16561-16570.

Research output: Contribution to journalArticle

Carven, GJ, Chitta, S, Hilgert, I, Rushe, MM, Baggio, RF, Palmer, M, Arenas, JE, Strominger, JL, Horejsi, V, Santambrogio, L & Stern, LJ 2004, 'Monoclonal Antibodies Specific for the Empty Conformation of HLA-DR1 Reveal Aspects of the Conformational Change Associated with Peptide Binding', Journal of Biological Chemistry, vol. 279, no. 16, pp. 16561-16570. https://doi.org/10.1074/jbc.M314315200
Carven, Gregory J. ; Chitta, Sriram ; Hilgert, Ivan ; Rushe, Mia M. ; Baggio, Rick F. ; Palmer, Michelle ; Arenas, Jaime E. ; Strominger, Jack L. ; Horejsi, Vaclav ; Santambrogio, Laura ; Stern, Lawrence J. / Monoclonal Antibodies Specific for the Empty Conformation of HLA-DR1 Reveal Aspects of the Conformational Change Associated with Peptide Binding. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 16. pp. 16561-16570.
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