Functional expression of a recombinant copper/zinc superoxide dismutase of filarial nematode, Brugia malayi.
A gene encoding a copper/zinc superoxide dismutase (Cu/ Zn-SOD) of a filarial nematode, Brugia malayi, has been isolated and the biochemical properties of a functionally expressed recombinant enzyme were investigated. The cloned complementary DNA contained a single open reading frame of 477 bp encoding 158 amino acids (aa), which conserved metal-binding residues as well as residues specific for Cu/Zn-SODs. Comparison of the deduced aa sequence of the enzyme with that of other helminthes species, including filarial worms, exhibited high degree of similarities (49-98%). Recombinant enzyme of 32 kDa had an isoelectric point of 6.6 and was shown to consist of 2 subunits linked by interchain disulfide bonds. Enzyme activity of the recombinant protein was inhibited by potassium cyanide and hydrogen peroxide but not by sodium azide. It showed a wide range of pH optima, i.e., 7.0-11.0 and was highly resistant to heat inactivation.
Lee
W G
WG
Department of Tropical and Endemic Parasitic Diseases, National Institute of Health, Seoul 122-701, Korea.
Hwang
J H
JH
Na
B K
BK
Cho
J H
JH
Lee
H W
HW
Cho
S H
SH
Kong
Y
Y
Song
C Y
CY
Kim
T S
TS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017178:Brugia malayi / Q000201:enzymology* / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015971:Gene Expression Regulation, Enzymologic; D005849:Gerbillinae; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D018510:RNA, Helminth / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D018510:RNA, Helminth; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013482:Superoxide Dismutase