DEVELOPMENT OF SURFACTANT HYDROPHOBIC PROTEIN 10,000

该建议旨在确定合成的结构和调节 表面活性剂相关疏水蛋白SAP-10 p。 这是哺乳动物表面活性剂的主要蛋白质组分。 该提案 提供了证据表明这是一种独特的脂质相关蛋白， 可能在表面活性剂的代谢和功能中发挥作用。 是 这种蛋白质可能是成功的围产期适应所必需的， 呼吸空气和缺乏表面活性剂与透明膜有关 疾病，婴儿发病率和死亡率增加的一个主要因素 早产儿 蛋白质将被分离，氨基酸组成和 从犬表面活性剂中确定的序列。 其功能测试 能够改变磷脂的性质、它们的吸收和 还将测试表面张力性质。 合成和分泌 将在35 S-蛋氨酸标记的II型中测定蛋白质 原代培养的上皮细胞。 抗血清和氨基酸序列将 用于筛选从犬产生的cDNA文库 在Lamdagt 11表达载体中的肺。 这种cDNA将被表征为 通过限制性酶切图谱和测序。 犬的cDNA 10，000道尔顿 蛋白质将被分离并用于筛选大鼠肺cDNA文库， 分离和鉴定该蛋白的大鼠cDNA。 10，000道尔顿合成控制的发育调控 将使用cDNA测定蛋白质以定量大鼠中的mRNA水平。 肺组织在妊娠后期，并通过测定合成和 分泌35 S-蛋氨酸标记的SAP-10 p的器官型培养物 胎鼠肺上皮细胞 这些研究将澄清起源， 10，000道尔顿疏水蛋白的结构和功能 表面活性剂，并导致进一步研究的分子基础， 围产期调节表面活性物质的合成和分泌。 这 为今后肺透明膜的治疗提供了有价值的信息 新生儿的疾病。

This proposal seeks to determine the structure and regulation of synthesis of surfactant associated hydrophobic protein of 10,000 daltons, SAP-10p. This is a major protein component of mammalian surfactant. The proposal provides evidence that this is a unique lipid associated protein which is likely to play a role in metabolism and function of surfactant. It is likely that this protein is required for successful perinatal adaptation to air breathing and lack of surfactant is associated with hyaline membrane disease, a major factor in the increased morbidity and mortality of infants born prematurely. The protein will be isolated, amino acid composition and sequence determined from canine surfactant. Functional testing of its ability to alter the properties of phospholipids, their absorption and surface tension properties will also be tested. Synthesis and secretion of the protein will be determined in 35S-methionine labelled Type II epithelial cells in primary culture. Antisera and amino acid sequence will be generated for use in screening of a cDNA library generated from canine lung in a Lamdagt11 expression vector. Such cDNA's will be characterized by restriction mapping and sequenced. cDNA's for the canine 10,000 dalton protein will be isolated and used to screen a rat lung cDNA library for isolation and characterization of the rat cDNA for this protein. Developmental regulation of control of synthesis of the 10,000 dalton protein will be determined using the cDNA to quantitate mRNA levels in rat lung tissue in late gestation and by determination of synthesis and secretion of 35S-methionine labelled SAP-10p by organotypic cultures of fetal rat lung epithelial cells. These studies will clarify the origin, structure and function of the 10,000 dalton hydrophobic protein in surfactant and lead to further study of the molecular basis for the perinatal regulation of surfactant synthesis and secretion. This information will be valuable in the future therapy of hyaline membrane disease in newborn infants.