The 4.5 Å Resolution Structure of a Bacterial Serine Protease from Streptomyces Griseus

作者：Penelope W. Codding, Louis T. J. Delbaere, Kota Hayakawa, W. L. B. Hutcheon, Michael N.G. James, Lubo Jurasek · 发表于：Canadian Journal of Biochemistry · 年份：1974 · DOI：10.1139/o74-034 · 被引用次数：19 · 研究领域：Peptidase Inhibition and Analysis、Enzyme Production and Characterization、Enzyme Structure and Function

Three crystalline modifications of the bacterial serine peptidase Streptomyces griseus Protease B have been grown. A 4.5 Å resolution electron density map of one form has been computed from the multiple isomorphous replacement (MIR) phases deduced from two heavy metal derivatives plus the anomalous dispersion effects of one of the derivatives. The crystalline modification used was grown from 0.7–1.0 M KH2PO4 at pH 4.2. These crystals have space group P21212 and unit cell dimensions of a = 44.15 (5) Å, b = 108.72 (10) Å, and c = 37.28 (5) Å. The crystal asymmetric unit contains a protein mass of approximately 19 000 daltons. The electron density map, mean figure of merit 0.80, clearly shows the molecular boundary; relatively long stretches of extended chain are discernable. The active site has been identified from a difference electron density map computed using the MIR protein phases and the amplitude differences between a crystal of the enzyme inhibited at the active serine in solution by p-iodobenzenesulfonyl fluoride (PIPSYL) and those from a crystal of the native enzyme. In addition to showing the site of the PIPSYL binding, there is an apparent conformational change in which the histidine side chain moves away from the serine residue by approximately 4.3 Å.