1. Susceptibility to HLA-DM Protein Is Determined by a Dynamic Conformation of Major Histocompatibility Complex Class II Molecule Bound with Peptide
- Author
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Sascha Lange, Marissa Bylsma, Marek Wieczorek, Abigail I Guce, Christian Freund, Lawrence J. Stern, Elizabeth D. Mellins, Liusong Yin, Wei Jiang, Peter Trenh, and Jana Sticht
- Subjects
Models, Molecular ,Conformational change ,Stereochemistry ,Immunology ,Molecular Sequence Data ,Antigen presentation ,Enzyme-Linked Immunosorbent Assay ,Peptide ,HLA-DM ,Crystallography, X-Ray ,Biochemistry ,Protein–protein interaction ,Epitopes ,Humans ,Amino Acid Sequence ,Surface plasmon resonance ,Binding site ,Nuclear Magnetic Resonance, Biomolecular ,Molecular Biology ,Peptide sequence ,chemistry.chemical_classification ,HLA-D Antigens ,Binding Sites ,Hydrogen Bonding ,Cell Biology ,Kinetics ,chemistry ,Peptides - Abstract
HLA-DM mediates the exchange of peptides loaded onto MHCII molecules during antigen presentation by a mechanism that remains unclear and controversial. Here, we investigated the sequence and structural determinants of HLA-DM interaction. Peptides interacting nonoptimally in the P1 pocket exhibited low MHCII binding affinity and kinetic instability and were highly susceptible to HLA-DM-mediated peptide exchange. These changes were accompanied by conformational alterations detected by surface plasmon resonance, SDS resistance assay, antibody binding assay, gel filtration, dynamic light scattering, small angle x-ray scattering, and NMR spectroscopy. Surprisingly, all of those changes could be reversed by substitution of the P9 pocket anchor residue. Moreover, MHCII mutations outside the P1 pocket and the HLA-DM interaction site increased HLA-DM susceptibility. These results indicate that a dynamic MHCII conformational determinant rather than P1 pocket occupancy is the key factor determining susceptibility to HLA-DM-mediated peptide exchange and provide a molecular mechanism for HLA-DM to efficiently target unstable MHCII-peptide complexes for editing and exchange those for more stable ones.
- Published
- 2014
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