title: Peptide-independent folding and CD8 alpha alpha binding by the nonclassical class I molecule, thymic leukemia antigen.
abstract: 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.
authors.affiliation: Department of Pathology, School of Medicine, Emory University, 1639 Pierce Drive, Atlanta, GA 30322, USA.
authors.forename: Dominique A
authors.initials: DA
authors.lastname: Weber
authors.forename: Antoine
authors.initials: A
authors.lastname: Attinger
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Kemball
authors.forename: Jerrod L
authors.initials: JL
authors.lastname: Wigal
authors.forename: Jan
authors.initials: J
authors.lastname: Pohl
authors.forename: Yi
authors.initials: Y
authors.lastname: Xiong
authors.forename: Ellis L
authors.initials: EL
authors.lastname: Reinherz
authors.forename: Hilde
authors.initials: H
authors.lastname: Cheroutre
authors.forename: Mitchell
authors.initials: M
authors.lastname: Kronenberg
authors.forename: Peter E
authors.initials: PE
authors.lastname: Jensen
mesh_terms: 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
chemical_list: 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