Purification of Taenia solium cysticerci superoxide dismutase and myoglobin copurification.
Superoxide dismutase from Taenia solium cysticerci ( Ts SOD) was purified by sequential ion exchange chromatography on quaternary-amino-ethyl-cellulose (QAE) followed by hydrophobic interaction on phenyl sepharose (PS) and chromatofocusing on a polybuffer exchanger 94 (PBE). Ts SOD is a 30 kDa molecular weight dimeric enzyme with 15 kDa monomers. It is partially negative, hydrophilic, with 6.3 isoelectric point and has 2,900 U/mg activity. Bovine erythrocyte SOD antibodies cross react with Ts SOD. This enzyme is 80% inhibited by 10 mM of KCN suggesting that it has a Cu/Zn active site. Furthermore, Ts SOD totally loses its activity at 100 degrees C for 4 min. The first 25 amino acids from the Ts SOD N-terminal are (M K A V X V M R G E E G V K G V V H F T Q A G D A). This sequence is 76% similar to the Schistosoma mansoni Cu/Zn SOD. By chance, myoglobin (Mb) was also found during the purification process. A 16 kDa band was recognized in immunoblotting by horse heart Mb antibodies in QAE, PS and PBE, the last-mentioned being found at pH 7.0. The first 15 amino acids from the amino terminal group (G L S D G E W Q L V L N V W G) in this 16 kDa protein are identical to several other Mbs which have been reported.
González
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R
Departamento de Microbiología y Parasitología, Facultad de Medicina, UNAM, Circuito interior, Ciudad Universitaria 04510 México, D. F. Mexico.
Mendoza-Hernández
G
G
Plancarte
A
A
D000595:Amino Acid Sequence; D000818:Animals; D003552:Cysticercus / Q000201:enzymology* / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification*; D011817:Rabbits; D020539:Sequence Analysis, Protein; D013482:Superoxide Dismutase / Q000302:isolation & purification*; D041201:Taenia solium / Q000201:enzymology* / Q000254:growth & development* / Q000378:metabolism
D003574:Cytochrome c Group; D003911:Dextrans; D009211:Myoglobin; C025614:sephadex; D013482:Superoxide Dismutase