Peptide-independent folding and CD8 alpha alpha binding by the nonclassical class I molecule, thymic leukemia antigen.
The nonclassical class I molecule, thymic leukemia (TL), has been shown to be expressed on intestinal epithelial cells and to interact with CD8(+) intraepithelial T lymphocytes. We generated recombinant soluble TL (T18(d)) H chains in bacteria as inclusion bodies and refolded them with beta(2)-microglobulin in the presence or absence of a random peptide library. Using a mAb, HD168, that recognizes a conformational epitope on native TL molecules, we observed that protein folds efficiently in the absence of peptide. Circular dichroism analysis demonstrated that TL molecules have structural features similar to classical class I molecules. Moreover, thermal denaturation experiments indicated that the melting temperature for peptide-free TL is similar to values reported previously for conventional class I-peptide complexes. Our results also show that CD8alphaalpha binding is not dependent on either TL-associated peptide or TL glycosylation.
Weber
Dominique A
DA
Department of Pathology, School of Medicine, Emory University, 1639 Pierce Drive, Atlanta, GA 30322, USA.
Attinger
Antoine
A
Kemball
Christopher C
CC
Wigal
Jerrod L
JL
Pohl
Jan
J
Xiong
Yi
Y
Reinherz
Ellis L
EL
Cheroutre
Hilde
H
Kronenberg
Mitchell
M
Jensen
Peter E
PE
D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques; D016827:CD8 Antigens / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D015789:HLA-A2 Antigen / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D017510:Protein Folding; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D013816:Thermodynamics; D001613:beta 2-Microglobulin / Q000378:metabolism
D000951:Antigens, Neoplasm; D016827:CD8 Antigens; C409469:CD8 antigen, alpha chain; D015789:HLA-A2 Antigen; D015395:Histocompatibility Antigens Class I; D008562:Membrane Glycoproteins; D019151:Peptide Library; D010455:Peptides; D001613:beta 2-Microglobulin; C028388:thymus-leukemia antigens