Optimization of Crystals of an Inhibitory Antibody of Urokinase Plasminogen Activator Receptor (uPAR) with Hydrogen Peroxide and Low Protein Concentration

Title: Optimization of Crystals of an Inhibitory Antibody of Urokinase Plasminogen Activator Receptor (uPAR) with Hydrogen Peroxide and Low Protein Concentration

Volume: 12 Issue: 7

Author(s): Yongdong Li, Xiaoli Shi, Graham Parry, Liqing Chen, Jennifer A. Callahan, Andrew P. Mazar and Mingdong Huang

Affiliation:

Keywords: hydrogen peroxide, crystal optimization, upar, antibody

Abstract: Optimization of protein crystal formation is often a necessary step leading to diffraction-quality crystals to enable collection of a full X-ray data set. Typical protein crystal optimization involves screening different components, e.g., pH, precipitants, and additives of the precipitant solution. Here we present an example using an inhibitory antibody of urokinase plasminogen activator receptor (uPAR) where such procedures did not yield diffracting crystals. In contrast, it was the treatment of the protein with hydrogen peroxide incubation and the protein concentration reduction that were found to be key factors in obtaining diffracting crystals. Final crystals diffracted to 1.75 Å, and belong to orthorhombic P212121 space group with unit cell parameters a=37.162 Å, b=84.474 Å, c=134.030 Å, and contain one molecule of Fab fragment of anti-uro kinase receptor antibody in the asymmetric unit.

Cite this article as:

Li Yongdong, Shi Xiaoli, Parry Graham, Chen Liqing, Callahan A. Jennifer, Mazar P. Andrew and Huang Mingdong, Optimization of Crystals of an Inhibitory Antibody of Urokinase Plasminogen Activator Receptor (uPAR) with Hydrogen Peroxide and Low Protein Concentration, Protein & Peptide Letters 2005; 12 (7) . https://dx.doi.org/10.2174/0929866054696073