Preparation of a protease resistant domain of the α-hemolysin monomer for crystallization

摘要: PS04. 03b. l3 PREPARATION OF A PROTEASE RESISTANT DOMAIN OF THE a-HEMOLYSIN MONOMER FOR CRYSTALLIZATION. Y.-D. Mo and JE Gouaux, Department of Biochemistry & Molecular Biology, University of Chicago, Chicago, IL a-Hemolysin (aHL) is secreted as a water-soluble monomer of 33 AleDa that self-assembles to fom1 heptameric transmembrane pores. Molecular entities along tlus pathway are tl1e water-soluble monomer (1), fuc membrane-bound monomer (2), the heptameric prepare (3) and the fully assembled heptameric channel (4) 1. To elucidate molecular mechanisms for the assembly, we aim to determine high resolution structures of each species using x-ray diffraction techniques. The goal of fuis work is to detemune tl1e structure of tl1e water-soluble monomer. However, fue crystallization of wild-type aHL in a monomeric, water soluble fom1 has proven-.. difficult. Therefore, we …