DEVELOPMENT OF SURFACTANT HYDROPHOBIC PROTEIN 10,000

这项建议旨在确定合成的结构和调节 表面活性物质相关疏水蛋白10,000道尔顿，SAP-10P。 这是哺乳动物表面活性物质的主要蛋白质成分。这项建议 提供证据表明这是一种独特的脂质相关蛋白，它是 可能对表面活性物质的代谢和功能起作用。它是 很可能这种蛋白质是围产期适应成功所必需的 呼吸空气和缺乏表面活性物质与透明膜有关 疾病，婴儿发病率和死亡率增加的主要因素 早产的早产的蛋白质将被分离，氨基酸组成和 由犬类表面活性物质确定的序列。智能交通系统的功能测试 能够改变磷脂的性质，它们的吸收和 还将测试表面张力特性。促性腺激素的合成和分泌 该蛋白质将在35S-蛋氨酸标记的II型中进行测定 原代培养的上皮细胞。抗血清和氨基酸序列将 可用于犬源基因文库的筛选 在Lamdagt11表达载体中构建肺脏表达载体。这样的cDNAs将被表征 通过限制性内切酶图谱和测序。犬10000道尔顿的cDNAs 将分离蛋白质并用于筛选大鼠肺cDNA文库 大鼠该蛋白基因的克隆与鉴定。 10,000道尔顿合成调控的发育规律 蛋白质将用该cDNA来定量测定大鼠的mRNA水平 妊娠晚期肺组织及肺组织合成和 ~(35)S-蛋氨酸标记SAP-10P在器官培养上的分泌 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 并为进一步研究表面活性剂的分子基础奠定了基础 围产期对表面活性物质合成和分泌的调节。这 这些信息将对未来透明膜的治疗有价值。 新生儿中的疾病。

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.