Characterization, molecular modelling and developmental expression of zebrafish manganese superoxide dismutase.
A 977 bp cDNA containing an open reading frame encoding 224 amino acid residues of manganese superoxide dismutase was cloned from zebrafish (zMn-SOD). The deduced amino acid sequence showed high identity with the sequences of Mn-SODs from human (85.1%) to nematode (61.6%). The 3-D structure model was superimposed on the relative domains of human Mn-SOD with the root mean square (rms) deviation of 0.0919 A. The recombinant mature zMn-SOD with enzyme activity was purified using His-tag technique. The half-life of the enzyme is approximately 48 min and its thermal inactivation rate constant k(d) is 0.0154 min(-1)at 70 degrees C. The enzyme was active under a broad pH (2.2-11.2) and in the presence of up to 4% SDS. Real-time RT-PCR assay was used to detect the zMn-SOD mRNA expression during the developmental stages following a challenge with paraquat. A high level expression of Mn-SOD mRNA was detected at the cleavage stage, but decreased significantly under paraquat treatment. The results indicated that Mn-SOD plays an important role during embryonic development.
Lin
Chi-Tsai
CT
Institute of Bioscience and Biotechnology & Center for Marine Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan, ROC.
Tseng
Wen-Chung
WC
Hsiao
Nai-Wan
NW
Chang
Hsiao-Huang
HH
Ken
Chuian-Fu
CF
D000595:Amino Acid Sequence; D000818:Animals; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000502:physiology; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
D013482:Superoxide Dismutase