title: Crystallization of the receptor-binding domain of parathyroid hormone-related protein in complex with a neutralizing monoclonal antibody Fab fragment.
abstract: 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 as a precipitant. The crystals belonged to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 72.6, b = 96.3, c = 88.5 A, and diffracted to 2.0 A resolution using synchrotron radiation. The crystal structure will shed light on the nature of the key residues of PTHrP that interact with the antibody and will provide insights into how the antibody is able to discriminate between PTHrP and the related molecule parathyroid homone.
authors.affiliation: Biota Structural Biology Laboratory, St Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia. bill.mckinstry@csiro.au
authors.forename: William J
authors.initials: WJ
authors.lastname: McKinstry
authors.forename: Galina
authors.initials: G
authors.lastname: Polekhina
authors.forename: Hannelore
authors.initials: H
authors.lastname: Diefenbach-Jagger
authors.forename: Koh
authors.initials: K
authors.lastname: Sato
authors.forename: Etsuro
authors.initials: E
authors.lastname: Onuma
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Gillespie
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Martin
authors.forename: Michael W
authors.initials: MW
authors.lastname: Parker