Biochemical Genetics of Carbonic Anhydrase Deficiencies

描述（由申请人提供）：本研究的广泛目标是研究碳酸酐酶基因家族的功能基因组学，以确定个体成员对健康和疾病的重要性。碳酸酐酶（carbonic anhydrases，CA）及其相关蛋白在呼吸、骨吸收、肾脏酸化、新生血管形成、信号转导、脑脊液和胃酸形成等多种生理过程中发挥重要作用。我们有五个具体目标： 1)对新发现的碳酸酐酶XIV的性质和功能基因组学进行表征。 2)表征CA IV敲除小鼠和CA IV/CA XIV双敲除的表型结果。 3)检验假设CA IV中的信号序列突变是显性遗传性视网膜色素变性（RP 17 q）的潜在缺陷。 4)描述线粒体碳酸酐酶CA VA和CA VB的特性和功能基因组学。 5)建立转基因小鼠模型以测试CA II/阴离子交换剂“代谢子”在整个动物中的功能重要性。 我们寻求新的支持，以一个计划，具有强大的生产力记录，因为我们最初发现的CA II缺乏综合征。这种疾病会影响骨骼、大脑和肾脏，是第一种与CA缺乏相关的疾病。我们将使用各种生物化学、细胞生物学、免疫学和分子遗传学方法。将通过靶向诱变产生个体CA缺陷的新型小鼠敲入和敲除小鼠模型。这些缺陷的功能后果将通过多个生理测量来表征。在适当情况下，将通过单缺陷小鼠之间的杂交产生多CA缺陷小鼠。这些研究应该加强我们对单个CA如何促进正常生理以及单CA和双CA缺乏如何产生疾病的理解。寻求的答案具有根本意义，并应提供信息，导致新的治疗方法CA缺陷。

DESCRIPTION (provided by applicant): The broad goal of this research is to study the functional genomics of the carbonic anhydrase gene family to determine the importance of individual members to health and disease. The sixteen known carbonic anhydrases (CAs) and CA-related proteins play important roles in diverse physiological processes including respiration, bone resorption, renal acidification, gluconeogenesis, signal transduction, and formation of cerebrospinal fluid and gastric acid. We have five specific aims: 1) Characterize the properties and functional genomics of newly discovered carbonic anhydrase XlV. 2) Characterize the CA IV knockout mouse and the phenotypic consequences of the CA IV/CA XIV double knockout. 3) Test the hypothesis that a signal sequence mutation in CA IV is the underlying defect in dominantly inherited retinitis pigmentosa (RP17q). 4) Characterize the properties and functional genomics of mitochondrial carbonic anhydrases CA VA and CA VB. 5) Create transgenic mouse models to test the functional importance of the CA I I/anion-exchanger "metabolon" in the whole animal. We seek renewed support for a program with a strong record of productivity since our initial discovery of the CA II deficiency syndrome. This disease affects bone, brain, and kidney and was the first disease associated with a CA deficiency. We will use a variety of biochemical, cell biological, immunological, and molecular genetic approaches. Novel mouse knock-in and knockout mouse models of individual CA deficiencies will be produced by targeted mutagenesis. Functional consequences of these deficiencies will be characterized by multiple physiological measurements. Mice deficient for multiple CAs will be produced, where appropriate, by intercrosses between singly deficient mice. These studies should enhance our understanding of how individual CAs contribute to normal physiology and how single and double CA deficiencies produce disease. The answers sought have fundamental significance, and should provide information leading to novel therapeutic approaches to CA deficiency.