Use of human surfactant low molecular weight apoproteins in the reconstitution of surfactant biologic activity.
Use of human surfactant low molecular weight apoproteins in the reconstitution of surfactant biologic activity.
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使用人表面活性剂低分子量脱辅基蛋白重建表面活性剂生物活性。
DOI：
10.1172/jci113391
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Cochrane,CG
中科院分区：
文献类型：
--
作者：
Revak,SD;Merritt,TA;Degryse,E;Stefani,L;Courtney,M;Hallman,M;Cochrane,CG
从人肺表面活性物质中分离得到两种低分子量脱辅基蛋白。SDS聚丙烯酰胺凝胶分析表明，一种蛋白质（SP 18）在未还原时具有18，000的表观分子量，还原后具有9，000 D。第二种蛋白质（SP 9）在存在或不存在还原剂的情况下在约9，000 D处迁移。这两种蛋白质都含有大量的疏水氨基酸。SP 18的NH 2-末端序列确定为：NH 2-phe-pro-ile-pro-leu-pro-tyr-。从成人肺cDNA文库中分离的cDNA克隆含有在内部位置编码人SP 18氨基末端序列的长开放阅读框。在脉动气泡表面计上评估磷脂（PL）与SP 9和SP 18的混合物降低表面张力的能力。加入1%的脱辅基蛋白导致15 s后的表面张力从单独PL的42.9 dyn/cm降低到含有SP 9和SP 18的制剂的16.7和6.3 dyn/cm。在胎兔中进行重构表面活性剂活性的体内评估。重组表面活性剂组成的PL +0.5%SP 18在分娩时气管内滴注导致肺顺应性显着增加，而0.5%SP 9的掺入产生了中度增加。这些数据显示了通过将分离的LMW脱辅基蛋白添加到定义的PL中来产生生物活性表面活性剂的能力。
Two low molecular weight (LMW) apoproteins were isolated from human pulmonary surfactant. SDS polyacrylamide gel analysis showed one protein (SP 18) to have an apparent molecular weight of 18,000 when unreduced and 9,000 D after reduction. The second protein (SP 9) migrated at approximately 9,000 D in the presence or absence of reducing agents. Both proteins contain a high number of hydrophobic amino acids. The NH2-terminal sequence of SP 18 was determined to be: NH2-phe-pro-ile-pro-leu-pro-tyr-. A cDNA clone isolated from a human adult lung cDNA library contained a long open reading frame encoding at an internal position the human SP 18 amino-terminal sequence. Mixtures of phospholipids (PL) and SP 9 and SP 18 were assessed for their capacity to reduce surface tensions on a pulsating bubble surfactometer. The addition of 1% apoprotein resulted in a reduction of surface tension after 15 s from 42.9 dyn/cm for PL alone to 16.7 and 6.3 dyn/cm for preparations containing SP 9 and SP 18, respectively. In vivo assessment of reconstituted surfactant activity was performed in fetal rabbits. Reconstituted surfactant consisting of PL + 0.5% SP 18 instilled intratracheally at delivery resulted in a marked increase in lung compliance, while the incorporation of 0.5% SP 9 yielded a moderate increase. These data show the ability to produce biologically active surfactant by the addition of isolated LMW apoproteins to defined PL.Images