COLLABORATION WITH PROF. ANDREAS PLUCKTHÜN INITIATED TO DEVELOP NMR-BASED METHODS FOR ELUCIDATION OF THE STRUCTURE OF GPCR-LIGAND COMPLEXES
14 July 2015, Leiden, NL
Recently, ZoBio and the laboratory of Prof. Andreas Pluckthün at the University of Zürich have started a collaboration funded through the Eurostars program. The primary goal of the project is to apply paramagnetic methods presently used within ZoBio to elucidate the structure of GPCR-ligand complexes. Currently, X-ray crystallography is the only structural method that is applied to GPCRs. However, two limitations to crystallography are routinely observed. First, the resolution of structures is often insufficient to observe or unambiguously place the ligand within the electron density. Second, capturing ligands that bind outside of the orthosteric site in crystals has proved extremely challenging.
ZoBio researchers have previously developed a method to triangulate the location of a ligand bound to a protein in collaboration with Prof. Marcellus Ubbink of Leiden University (Guan et al., JACS, 2013, 135(15): 5859-5868). The technique makes use of the distance and orientation-dependent effects of a paramagnetic Lanthanide (Ln) ion attached at selected sites on the surface of a protein using so-called CLanPs. The project will utilize the GPCR stabilizing CHESS technology developed in Prof. Pluckthün’s laboratory. The ability to express these highly stable GPCRs in E. coli will enable rapid engineering of the CLanP attachment sites. The resulting paramagnetic constraints will enable the structure elucidation of allosteric modulators that have been discovered using ZoBio’s expertise in fragment based drug discovery. Such a structure would be a world’s first and could provide insight into the allosteric modulation of GPCRs by small molecules.