MOLECULAR-CLONING AND FUNCTIONAL EXPRESSION OF A CDNA FOR RAT PHOSPHOLIPID HYDROPEROXIDE GLUTATHIONE-PEROXIDASE - 3'-UNTRANSLATED REGION OF THE GENE IS NECESSARY FOR FUNCTIONAL EXPRESSION

被引:59
作者
IMAI, H
SUMI, D
HANAMOTO, A
ARAI, M
SUGIYAMA, A
CHIBA, N
KUCHINO, Y
NAKAGAWA, Y
机构
[1] KITASATO UNIV,SCH PHARMACEUT SCI,MINATO KU,TOKYO 108,JAPAN
[2] NATL INST CANC RES,DEPT BIOPHYS,CHUO KU,TOKYO 104,JAPAN
[3] JAPAN ENERGY CORP,TODA,SAITAMA 335,JAPAN
关键词
CDNA; EXPRESSION; PHOSPHOLIPID HYDROPEROXIDE GLUTATHIONE PEROXIDASE; SELENOCYSTEINE; 3'-UNTRANSLATED REGION;
D O I
10.1093/jb/118.5.1061
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A full-length cDNA clone for phospholipid hydroperoxide glutathione peroxidase (PHGPx) was isolated from a rat brain. The cDNA was 0.761 kb in length and encoded 170 amino acids, which included a TGA-encoded selenocysteine at residue 46. The protein has a calculated molecular mass of 19,473 Da. We succeeded in the transient functional expression of a full-length cDNA for PHGPx, which includes the 3'-UTR, in COS-7 cells at the first attempt. Deletion of the 3'-UTR prevented the expression of the PHGPx activity and the incorporation of [Se-75]selenious acid into the monomeric 19.7 kDa PHGPx protein, Thus, the 3'-UTR of the cDNA for PHGPx was required for the functional expression of PHGPx. Northern blot analysis demonstrated that the mRNA for PHGPx was widely expressed in normal rat tissues, especially in the testis, The mRNA levels of PHGPx in the cultured cells such as hepatomas, neuronal cells, nephroblastoma, and mammary myo-epithelial cells were higher than those of the tissues. The ratio of PHGPx to cytosolic glutathione peroxidase (cGPx) was significantly high in the testis and relatively high in the cultured cells. The mRNA levels of PHGPx in tissues were lower than those of cGPx.
引用
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页码:1061 / 1067
页数:7
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