CATALYTIC MECHANISM AND INHIBITION OF GOLGI ALPHA-MANNOSIDASE II

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We previously determined the structure of the glycosylation enzyme alpha-mannosidase II, a glycosyl hydrolase of the Family 38 family targetted by potential anti-metastatic compounds. Largely with data from CHESS, we have been analyzing the atomic basis of catalysis and inhibition of the enzyme through structures of complexes with intermediates and several classes of inhibitors. Recently, we performed two high-throughput screens of chemical libraries looking for novel inhibitors of the enzyme. This resulted in 10-12 hits of varying affinity and properties. We are interested in examining how these bind to the structure. In particular, some may be non-competitive inhibitors and may occupy sites distant from the catalytic centre, which will be of interest for future inhibitor design. We would like to use synchrotron radiation for two promary reasons. Firstly, we can get a data set in minutes, rather than hours, which will let us quickly examine several of these complexes in a short time. Secondly, we can get almost atomic resolution (1.2-1.4A) data from these crystals at CHESS. This resolution can be critical in understanding conformational distortions and determining precise interatomic distances.