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. BChE is a potential drug for two contemporary wars. BChE could serve as a harmless detoxifier of cocaine and has the potential of protecting troops and first-responders from the harmful and long-term effects of nerve-agents. The long-range goal is to produce kilogram quantities of human BChE for therapeutic use in humans. In order to be therapeutically useful, BChE must have a long residence time in the circulation, which means it must be in tetrameric form. Towards these ends, we propose a research study that will elucidate the structural basis of tetramer formation. The principal technique to be employed is macromolecular crystallography. There are no reports of crystals of full-length recombinant BChE.Thus far we have successfully crystallized monomeric full length BChE.However, the data collected on the local low intensity rotating anode X-ray source is only complete to 4.5 Angstroms. Attempts are underway to determine the structure of monomeric full-length BChE by molecular replacement using truncated BChE as the search model, thus far the low resolution difference and double difference electron density maps generated shows density into which a poly-alanine helix representing the last 45 amino-acids can be placed. The low resolution of the data limits the quality of our structure. It is likely that better data to higher resolution can be collected at a higher intensity X-ray source such as a synchrotron.
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