Crystal structures of γ-glutamyltranspeptidase in complex with inhibitors

γ-Glutamyltranspeptidase (GGT; EC 2.3.2.2) is involved in the degradation of γ-glutamyl compounds such as glutathione (GSH; γ-glutamyl-cysteinyl-glycine) . A major physiological role of this enzyme is to cleave the extracellular GSH as a source of cysteine for intracellular glutathione biosynthesis. Another crucial role of GGT is to cleave glutathione-S-conjugates as a key step in detoxification of xenobiotics and drug metabolism. In mammals, GGT has been implicated in physiological disorders such as Parkinson's disease, other neurodegenerative diseases including Alzheimer's disease and cardiovascular disease. The indispensable roles played by GGT in GSH-mediated detoxification and cellular response to oxidative stress suggest that GGT is an attractive pharmaceutical target. We here report the binding mode of acivicin, a well-known glutamine antagonist, to B. subtilis GGT at 1.8 Å resolution showing that acivicin is bound to the Oγ atom of Thr403, the catalytic nucleophile of the enzyme, through its C3 atom [1]. The observed electron density around the C3 atom was best fitted to the planar and sp2 hybridized carbon, consistent with a simple nucleophilic substitution of Cl at the imino carbon by Oγ atom of Thr403. Furthermore, comparison of three bacterial enzymes, the GGTs from E. coli, H. pylori and B. subtilis in complex with acivicin, showed significant diversity in the orientation of the dihydroisoxazole ring among three GGTs. The differences are discussed in terms of the recognition of the α-amino and α-carboxy groups in preference to the dihydroisoxazole ring as observed in time-lapse soaking crystal structures of B. subtilis GGT with acivicin.

Particle statistics in powder diffraction method

The statistical variation in observed powder diffraction intensity is often dominated by the limited number of crystallites that satisfy the diffraction condition. This effect has been termed with particle statistics in powder diffraction method [1]. We have reported experimental validation of the theory for particle statistics about symmetric reflection mode measurements, and proposed a new method to evaluate the crystallite size and its distribution by statistical analysis based on a generalized theoretical framework [2]. We have also demostrated improvement of structure refinement by application of the maximum likelihood estimation to statistical model explicitly taking the errors caused by the particle statistics into account [3]. As the formula suitable for the particle statistics about a continuously spinning flat specimen is still unclear, three kinds of diffraction intensity measurements, (1) in-plane (Φ)-scan, rocking angle (Ω)-scans for (2) stationary and (3) spinning flat specimens, have been conducted for standard powder samples of Si (NIST SRM640c) and lanthanum hexaboride (NIST SRM660a) at a synchrotron powder diffraction beamline BL-4B2 at the Photon Factory, KEK in Tsukuba. The statistical variance observed inΦ-scan and Ω-scan measurements for stationary specimens have shown cosecant dependence of the effective number of diffracting crystallites, just as predicted by the traditional theory. The statistical variance in Ω-scan measurements for spinning specimens has also shown a cosecant behaivior rather than the squared cosecant dependence suggested by de Wolff [1].