Partial nuclear magnetic resonance assignments of copper zinc superoxide dismutase from Brucella abortus
Date
1993
Authors
Chen, Yen-Liang
Major Professor
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Kintanar, Agustin
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Abstract
The ulitmate goal of this investigation is to study the structure-function relationships of Brucella abort-us copper zinc superoxide dismuta.se (BA Cu-Zn SOD). B. abortus is a pathogenic bacterium which infects cattle and causes abortion. Control of brucellosis is a significant cost to the cattle industry. In addition, humans can beinfected by this pathogen and the infection is potentially lethal. The use of SOD to control superoxide radical concentration has been hotly pursued and we hope that by understanding its three dimensional structure, we may contribute to the design of a better SOD for therapeutic purposes. In this investigation, cost-effective labelling of B. abortus Cu-Zn SOD was achieved using molecular cloning techniques. In addition, we have successfully reproduced the purification procedure of B. abortus cloned in E. coli done by Bricker (Bricker et al., 1990) with minor alterations in the procedure. Also, we have reproduced the reconstitution procedure used in bovine Cu-Zn SOD for B. abortus Cu-Zn SOD with minor alterations in the procedure (Beem et al., 1974). Furthermore, we have assigned almost all the histidine imidazole protons in the active site. Last of all, we are still working on the most time consuming phase of our investigation, the sequential assignment of B. abortus Cu-Zn SOD. Currently, approximately 50 residues have been assigned.
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