Purification and properties of Cu-Zn superoxide dismutase extracted from Brucella abortus strain 19
- ARS-USDA, Ames, IA (United States)
Recent work showed that a recombinant 20 kDa protein from Brucella abortus expressed in E. coli is a Cu-Zn superoxide dismutase (SOD). Western blot and ELISA results indicated that cattle with brucellosis have antibody to SOD. Here the authors report the purification and properties of the native B. abortus Cu-Zn SOD. SOD was extracted from methanol-killed Brucella abortus strain 19 with 0.1 M sodium citrate-1.0 M sodium chloride solution. The extract was dialyzed and protein precipitated by ammonium sulfate at 70-100% saturation was collected. The SOD was purified by HPLC anion exchange chromatography. SOD activity was assayed with a coupled enzyme assay using xanthine oxidase-cytochrome C reduction assay. The authors determined that the Brucella SOD is present in two molecular forms both inhibitable with KCN with Ki's of 0.32 mM and 4.98 mM, respectively. No other form of SOD was identified in the extract. Polyclonal antibody to SOD and polyclonal antibody to SOD synthetic peptide residues 134-143 inhibited SOD activity by 50% and 13%, respectively. Both SOD and the synthetic peptide inhibited binding of anti-SOD antibody to SOD by 60% and 20%, respectively. Based on these results the SOD and its amphipathic peptide will be considered as candidates for the design of synthetic multiple peptide vaccines and diagnostic reagents for bovine brucellosis.
- OSTI ID:
- 5372888
- Report Number(s):
- CONF-9104107-; CODEN: FAJOE
- Journal Information:
- FASEB Journal (Federation of American Societies for Experimental Biology); (United States), Vol. 5:4; Conference: 75. annual meeting of the Federation of American Societies for Experimental Biology (FASEB), Atlanta, GA (United States), 21-25 Apr 1991; ISSN 0892-6638
- Country of Publication:
- United States
- Language:
- English
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COPPER
METABOLISM
SUPEROXIDE DISMUTASE
CHEMICAL PROPERTIES
ZINC
BRUCELLA
ENZYME IMMUNOASSAY
ESCHERICHIA COLI
FRACTIONATION
ION EXCHANGE CHROMATOGRAPHY
LIQUID COLUMN CHROMATOGRAPHY
BACTERIA
BIOASSAY
CHROMATOGRAPHY
ELEMENTS
ENZYMES
IMMUNOASSAY
METALS
MICROORGANISMS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PROTEINS
SEPARATION PROCESSES
TRANSITION ELEMENTS
560300* - Chemicals Metabolism & Toxicology