Crystallization of the receptor-binding domain of parathyroid hormone-related protein in complex with a neutralizing monoclonal antibody Fab fragment
William J McKinstry, Galina Polekhina, Hannelore Diefenbach-Jagger, Koh Sato, Etsuro Onuma, Matthew T Gillespie, Thomas J Martin, Michael W Parker
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | INT UNION CRYSTALLOGRAPHY | Published : 2009
Parathyroid hormone-related protein (PTHrP) plays an important role in regulating embryonic skeletal development and is abnormally regulated in the pathogenesis of skeletal complications observed with many cancers and osteoporosis. It exerts its action through binding to a G-protein-coupled seven-transmembrane cell-surface receptor (GPCR). Structurally, GPCRs are very difficult to study by X-ray crystallography. In this study, a monoclonal antibody Fab fragment which recognizes the same region of PTHrP as its receptor, PTH1R, was used to aid in the crystallization of PTHrP. The resultant protein complex was crystallized using the hanging-drop vapour-diffusion method with polyethylene glycol ..View full abstract
This work was supported in part by grants from the Cancer Council of Victoria and the National Health and Medical Research Council of Australia (NHMRC), with infrastructural support as the Australian Cancer Research Foundation Rational Drug Discovery Facility. This work, including the use of the BioCARS sector, was also supported by the Australian Synchrotron Research Program, which is funded by the Commonwealth of Australia under the Major National Research Facilities Program. Use of the Advanced Photon Source was supported by the US Department of Energy, Basic Energy Sciences, Office of Energy Research. We thank Dr Harry Tong and other staff at BioCARS for their help with data collection during our visit to the Advanced Photon Source. WJM was an NHMRC Industry Fellow, GP is an R. D. Wright NHMRC Fellow, MTG is an NHMRC Principal Research Fellow and MWP is an Australian Research Council Federation Fellow and an NHMRC Honorary Fellow.