Molecular Basis of Natural Killer Cell Tumor Target Recognition: The NKr/MHC Class I Complex
Abstract
We have proposed to characterize at a molecular level the recognition determinants of stimulatory and inhibitory NK receptors and the class I molecules they bind. We have pursued this via the heterologous expression of wild type and mutant NK receptors for the purpose of the determination of the atomic structure of an NK receptor/ class I MHC complex via X-ray crystallography. We have investigated several methods for the over expression of the murine inhibitory NK receptor Ly49c. The results of this survey indicate that the most tractable solution is the over-expression of Ly49c in E. coli, where the protein is found in an insoluble inclusion body fraction. We have developed a protocol for the in vitro refolding of denatured Ly49c from this inclusion body material. This work now allows us to investigate, by in vitro killing assay and/or direct biochemical assays, the nature of the interaction between Ly49c and its ligand, H2-K(b). We have also begun screening for crystallization conditions with refolded Ly49c, for the purpose of determining its atomic structure.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 1999
- Accession Number
- ADA385884
Entities
People
- Charles A. Hasemann
Organizations
- University of Texas at Dallas