Role of chain pairing for the production of functional soluble IA major histocompatibility complex class II molecules
| Autor: | K C Garcia, Luc Teyton, C.A. Scott, Ian A. Wilson, Francis R. Carbone |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Leucine zipper
DNA Complementary T-Lymphocytes Immunology Molecular Sequence Data Receptors Antigen T-Cell Biology Major histocompatibility complex Transfection Polymerase Chain Reaction Cell Line Mice Antigen HLA-DQ Antigens MHC class I Immunology and Allergy Animals Humans Amino Acid Sequence Peptide sequence Cells Cultured DNA Primers chemistry.chemical_classification Leucine Zippers HLA-DQ Antigen Histocompatibility Antigens Class II Thrombin Articles Molecular biology Recombinant Proteins Cell biology Histocompatibility Kinetics Drosophila melanogaster chemistry biology.protein Glycoprotein |
| Zdroj: | The Journal of Experimental Medicine |
| ISSN: | 0022-1007 |
| Popis: | Structural studies of cellular receptor molecules involved in immune recognition require the production of large quantities of the extracellular domains of these glycoproteins. The murine major histocompatibility complex (MHC) class II-restricted response has been extensively studied by functional means, but the engineering and purification of the native, empty form of the most-studied murine MHC class II molecule, IA, has been difficult to achieve. IA molecules, which are the murine equivalent of human histocompatibility leukocyte antigen-DQ molecules, have a low efficiency of chain pairing, which results in poor transport to the cell surface and in the appearance of mixed isotype pairs. We have engineered soluble IA molecules whose pairing has been forced by the addition of leucine zipper peptide dimers at their COOH-terminus. The molecules are secreted "empty" into the extracellular medium and can be loaded with single peptide after purification. These IA molecules have been expressed in milligram quantity for crystallization as well as for activation of T cells and measurement of MHC class II-T cell receptor interactions. |
| Databáze: | OpenAIRE |
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