Crystallization and preliminary X-ray crystallographic analysis of the protease inhibitor ecotin in complex with chymotrypsin

Ecotin, a homodimeric protein composed of 142-residue subunits, is a novel protease inhibitor present in the periplasm of Escherichia coli. It shows a broad inhibitory specificity towards a group of serine proteases and binds two molecules of protease to form a tetrameric complex in a distinct chelation mechanism. The ecotin-chymotrypsin complex has been crystallized in the triclinic space group P1 with unit-cell parameters a = 57.29, b = 57.39, c = 79.75 Angstrom, alpha = 91.49, beta = 88.63 and gamma = 112.45 degrees. The asymmetric unit contains the whole tetrameric complex, consisting of two molecules of chymotrypsin bound to the ecotin dimer, with a corresponding crystal volume per protein mass (V-M) of 2.58 Angstrom(3) Da(-1) and a solvent fraction of 48.9%. The crystals diffract beyond 2.0 Angstrom with Cu K alpha X-rays and are very stable in the X-ray beam. Native X-ray data have been collected from a crystal to approximately 2.0 Angstrom Bragg spacing.