Endozepine is a mitochondrial-associated protein that plays a role in ceramide-induced apoptosis, possibly by modulating the release of cytochrome C from mitochondria. (For more biological details, see Dr Mervin Meyer, UWC, or Jasper.) It shares significant homology with Acyl-CoA-binding proteins, and preliminary data suggests that it may bind directly to ceramide. Endozepine has a molecular weight of approximately 10kDa, which makes it suitable for structure determination using either NMR or X-ray.
The protein has been recombinantly expressed as a GST fusion and much of the optimisation of expression and solubility has been done. Preliminary NMR spectra show it to be well-folded. The project will involve expression of 15N and 13C labelled samples and generation of full backbone assignments using NMR. These will be used in conjunction with homology models to map the binding site of ceramide and other potential ligands and finally the structure of Endozepine will be determined in collaboration with David.