DM loss in k haplotype mice reveals isotype-specific chaperone requirements.

Journal article

DM actions as a class II chaperone promote capture of diverse peptides inside the endocytic compartment(s). DM mutant cells studied to date express class II bound by class II-associated invariant chain-derived peptide (CLIP), a short proteolytic fragment of the invariant chain, and exhibit defective peptide-loading abilities. To evaluate DM functional contributions in k haplotype mice, we engineered a novel mutation at the DMa locus via embryonic stem cell technology. The present experiments demonstrate short-lived A(k)/CLIP complexes, decreased A(k) surface expression, and enhanced A(k) peptide binding activities. Thus, we conclude that DM loss in k haplotype mice creates a substantial pool of empty or loosely occupied A(k) conformers. On the other hand, the mutation hardly affects E(k) activities. The appearance of mature compact E(k) dimers, near normal surface expression, and efficient Ag presentation capabilities strengthen the evidence for isotype-specific DM requirements. In contrast to DM mutants described previously, partial occupancy by wild-type ligands is sufficient to eliminate antiself reactivity. Mass spectrometry profiles reveal A(k)/CLIP and a heterogeneous collection of relatively short peptides bound to E(k) molecules. These experiments demonstrate that DM has distinct roles depending on its specific class II partners.

Authors: Koonce, C; Wutz, G; Robertson, E; Vogt, AB; Kropshofer, H

• Publication status: Published
• Journal: Journal of Immunology
• Volume: 170
• Issue: 7
• Pages: 3751-3761
• Publication date: 2003-04-01
• DOI: 10.4049/jimmunol.170.7.3751
• EISSN: 1550-6606
• ISSN: 0022-1767
• Language: English
• Keywords: Mice, Inbred BALB C; Antigens, Differentiation, B-Lymphocyte; Male; Mice, Inbred CBA; H-2 Antigens; Antigen Presentation; Crosses, Genetic; Muramidase; RNA Editing; Mice, Inbred C57BL; Haplotypes; Histocompatibility Antigens Class II; Gene Deletion; Mice; Molecular Sequence Data; Female; Molecular Chaperones; Mice, Knockout; Amino Acid Sequence; Animals; Self Tolerance; Peptide Fragments