NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase: immunochemical characterization of the lung enzyme from pregnant rabbits.
NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase: immunochemical characterization of the lung enzyme from pregnant rabbits.
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NAD（）依赖性15-羟基前列腺素脱氢酶：怀孕兔肺酶的免疫化学特征。
DOI：
10.1016/0003-9861(90)90488-k
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发表时间：
1990
影响因子：
3.9
通讯作者：
Bergholte,JM
中科院分区：
文献类型：
--
作者：
Okita,RT;Sinning,AR;Okita,JR;Tai,HH;Markwald,RR;Bergholte,JM
从孕兔肺组织中纯化了NAD+依赖的15-羟前列腺素脱氢酶（PGDH），并制备了抗PGDH的多克隆抗体。Westernblot分析表明，该抗体在105，000 g孕兔肺组织上清中鉴定的蛋白质分子量为30 kDa，与纯化的PGDH共迁移。妊娠兔（妊娠25- 28天）肺PGDH的比活性为36.7 nmol NADH形成/min/mg蛋白，而非妊娠兔为0.3 nmol NADH形成/min/mg蛋白。尽管抗肺PGDH抗体可抑制未孕兔肺胞液中的PGDH活性，但Western印迹分析未检测到30-kDa蛋白。在妊娠期间对该30-kDa蛋白质的检查表明，该蛋白质在10天后存在，并且该蛋白质的量从第10天至第28天增加。免疫化学反应蛋白的这种增加与PGDH比活性在10至28天之间的显著增加相关。在妊娠25 ~ 28天的家兔卵巢中也观察到一种免疫化学反应蛋白，其PGDH比活性为19.3 nmol NADH形成/min/mg蛋白。在未孕兔卵巢中仅检测到微量PGDH活性。抗兔肺PGDH在妊娠或非妊娠兔的脑、肾、膀胱、子宫、肝和心脏组织中未检测到30 kDa蛋白。当用抗兔肺PGDH检查兔或人胎盘胞质溶胶时，通过Western印迹分析仅观察到微弱的30-kDa条带。制备的抗人胎盘PGDH单克隆抗体不识别孕兔肺中的30 kDa条带。定位研究表明，妊娠兔肺上皮细胞的免疫化学染色显着增加，与非妊娠兔相比。肺上皮细胞而不是内皮细胞被鉴定为含有PGDH。
A polyclonal antibody was produced in guinea pig against the lung NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) purified from pregnant rabbits. Western blot analysis demonstrated that the protein identified by this antibody in the 105,000gsupernatant fraction of lung tissue from pregnant rabbits had a molecular mass of 30 kDa and comigrated with the purified PGDH. The specific activity of the lung PGDH in pregnant rabbits (25- to 28-day gestations) was 36.7 nmol NADH formed/min/mg protein compared to 0.3 nmol NADH formed/min/mg protein in nonpregnant rabbits. Although the PGDH activity in the lung cytosol of nonpregnant rabbits was inhibited by the anti-lung PGDH antibody, the 30-kDa protein was not detected by Western blot analysis. An examination of this 30-kDa protein during the gestational period indicated that the protein was present after 10 days and the amount of the protein increased from Day 10 to Day 28. This increase in the immunochemically reactive protein correlated with the marked increase in PGDH specific activity between 10 and 28 days. An immunochemically reactive protein also was observed in the ovary of 25- to 28-day pregnant rabbits and the specific activity of the ovary PGDH was 19.3 nmol NADH formed/min/mg protein. Only trace levels of the PGDH activity were detected in the ovaries of nonpregnant rabbits. A 30-kDa protein was not detected by the anti-rabbit lung PGDH in brain, kidney, bladder, uterus, liver, and heart tissue of pregnant or nonpregnant rabbits. When rabbit or human placental cytosol was examined with the anti-rabbit lung PGDH only faint 30-kDa bands were observed by Western blot analysis. A monoclonal antibody prepared against human placental PGDH did not recognize the 30-kDa band in the pregnant rabbit lung. Localization studies indicated a marked increase in immunochemical staining in pulmonary epithelial cells of pregnant rabbits as compared to nonpregnant rabbits. Lung epithelial cells but not endothelial cells were identified as containing the PGDH.
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DOI：
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发表时间：
1971
期刊：
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作者：
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DOI：
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发表时间：
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期刊：
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