Enzymes of Skin Lipid Metabolism

皮肤脂质代谢酶

• 批准号: 6982985
• 负责人: DAVID W. RUSSELL
• 金额: $ 34.32万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6982985
• 关键词: CHO cells; acyltransferase; cell differentiation; cholesterol esters; enzyme activity; enzyme induction /repression; fibroblasts; genetic regulation; genetically modified animals; keratinocyte; laboratory mouse; lipid metabolism; phenotype; polymerase chain reaction; protein localization; skin; sulfotransferase

DESCRIPTION (provided by applicant): Cholesterol sulfate and wax monoesters are abundant lipids in the skin whose biological roles and biosynthetic pathways remain largely undefined. Cholesterol sulfate is synthesized by the SULT2B1 sulfotransferase enzyme and is postulated to be an essential regulatory and structural component in the formation of the epidermal lipid barrier. The biosynthetic pathway that produces wax monoesters has not been defined in mammals but by analogy to plants and lower organisms is thought to involve the sequential actions of two enzymes, a fatty acyl coenzyme A (CoA) reductase and a wax ester synthase. Our initial goal will be to characterize the phenotype of mice that lack cholesterol sulfotransferase. Targeted gene disruption has produced SULT2B1 knockout mice that will be studied with respect to skin barrier function, lipid metabolism and reproduction. The second goal is to determine the role of cholesterol sulfotransferase in vitro by characterizing Chinese hamster ovary cell lines that over-express SULT2B1 and primary mouse embryo fibroblasts and keratinocytes that lack this enzyme. Sterol metabolism, gene regulation, and differentiation of these cells will be analyzed. Our third goal is to define the mammalian wax monoester biosynthetic pathway. Bioinformatics and expression cloning have produced mouse cDNAs encoding two fatty acyl CoA reductases and a wax synthase. The biochemical properties, subcellular Iocalizations, and tissue distributions of these enzymes will be determined. The final objective is to produce mice that are deficient in the wax synthase enzyme. The resulting knockout animals will be used to determine the function of wax monoesters in the skin, eye, and immune system. The proposed studies will provide new insight into the enzymes of skin lipid metabolism and the roles that unique lipids play in preventing infection, dehydration, and abrasion. The research is relevant to skin disease, dry eye syndrome, and cholesterol metabolism.

描述（由申请人提供）： 硫酸胆固醇和蜡单酯是皮肤中丰富的脂质，其生物学作用和生物合成途径在很大程度上仍不清楚。硫酸胆固醇由 SULT2B1 磺基转移酶合成，被认为是表皮脂质屏障形成中的重要调节和结构成分。产生蜡单酯的生物合成途径尚未在哺乳动物中得到定义，但通过与植物和低等生物体的类比，人们认为涉及两种酶的连续作用，即脂肪酰辅酶 A (CoA) 还原酶和蜡酯合酶。我们的最初目标是表征缺乏胆固醇磺基转移酶的小鼠的表型。靶向基因破坏已产生 SULT2B1 基因敲除小鼠，将对其皮肤屏障功能、脂质代谢和繁殖进行研究。第二个目标是通过表征过度表达 SULT2B1 的中国仓鼠卵巢细胞系以及缺乏这种酶的原代小鼠胚胎成纤维细胞和角质形成细胞，确定体外胆固醇磺基转移酶的作用。将分析这些细胞的甾醇代谢、基因调控和分化。我们的第三个目标是定义哺乳动物蜡单酯生物合成途径。生物信息学和表达克隆已经产生了编码两种脂肪酰辅酶A还原酶和一种蜡合酶的小鼠cDNA。这些酶的生化特性、亚细胞定位和组织分布将被确定。最终目标是培育出缺乏蜡合酶的小鼠。由此产生的基因敲除动物将用于确定蜡单酯在皮肤、眼睛和免疫系统中的功能。拟议的研究将为皮肤脂质代谢酶以及独特脂质在预防感染、脱水和擦伤中的作用提供新的见解。该研究与皮肤病、干眼症和胆固醇代谢有关。