Graduate School of Science and Technology, Nagasaki University, Nagasaki 852-8521, Japan.
Comp Biochem Physiol B Biochem Mol Biol. 2011 Apr;158(4):289-96. doi: 10.1016/j.cbpb.2010.12.007. Epub 2011 Jan 1.
Manganese-superoxide dismutase (Mn-SOD) from Japanese flounder (Paralichthys olivaceus) hepatopancreas has been purified with high purification (781-fold) and recovery (10.8%). The molecular mass of the purified enzyme was estimated to be 26kDa by SDS-PAGE under reducing conditions. In activity staining by native-PAGE, the Japanese flounder Mn-SOD gave three active bands and exhibited KCN-insensitive activity. In addition, the electrophoretic mobility of this enzyme was observed to be faster than that of Japanese flounder Cu,Zn-SOD. On the other hand, the N-terminal amino acid sequence of this Mn-SOD was determined to be 16 amino acid residues, and the sequence showed high homology to other Mn-SODs but not Japanese flounder Cu,Zn-SOD. Analysis of nucleotide and deduced amino acid sequences revealed that the Mn-SOD cDNA consisted of a 64bp 5'-non-coding region, a 675bp open reading frame encoding 225 amino acids, and a 465bp 3'-non-coding region. The first 27 amino acids containing a mitochondria-targeting signal were highly conserved among other Mn-SODs.
从日本牙鲆（Paralichthys olivaceus）肝胰腺中纯化了锰超氧化物歧化酶（Mn-SOD），其具有高纯度（781 倍）和回收率（10.8%）。通过还原条件下的 SDS-PAGE，估计纯化酶的分子量为 26kDa。在天然-PAGE 的活性染色中，日本牙鲆 Mn-SOD 给出了三条活性带，并表现出对 KCN 的不敏感活性。此外，该酶的电泳迁移率被观察到比日本牙鲆 Cu，Zn-SOD 更快。另一方面，该 Mn-SOD 的 N-末端氨基酸序列被确定为 16 个氨基酸残基，该序列与其他 Mn-SOD 具有高度同源性，但与日本牙鲆 Cu，Zn-SOD 不同。核苷酸和推导的氨基酸序列分析表明，Mn-SOD cDNA 由 64bp 的 5'-非编码区、675bp 的开放阅读框编码 225 个氨基酸和 465bp 的 3'-非编码区组成。包含线粒体靶向信号的前 27 个氨基酸在其他 Mn-SOD 中高度保守。
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