Functional and biochemical studies of ADAM19

性状（由申请人提供）：金属蛋白酶-去整合素（ADAM）是一种 膜锚定糖蛋白家族，在受精中起关键作用， 在神经发生中，以及在发育过程中EGF受体配体的活化中。的 这项建议的主要目标是揭示围产期的潜在机制， 缺乏广泛表达的ADAM 19的小鼠的致死表型， 评估ADAM 19在蛋白质胞外域脱落中的预测作用。 蛋白质胞外域脱落导致多种结构上的 和功能多样的蛋白质从质膜，并最近 作为一种重要的翻译后调节机制， 释放的蛋白质。胞外域脱落对于正确的信号传导是必不可少的 通过EGF受体配体，Notch介导的侧向抑制，限制TNF 受体介导的炎症反应，并调节几个方面， 轴突导向因为第一个确定的脱落酶，TNF α 转化酶是ADAM，并且因为IIMP抑制研究排除了目前的 已知基质金属蛋白酶（MMP）作为大多数蛋白质的脱落酶，ADAM具有 成为最佳候选胞外域脱落酶。我们的假设是 缺乏ADAM 19的小鼠的表型至少部分是由于 在一种或多种蛋白质的胞外域脱落中。具体目标是 项目有： 1)对缺乏ADAM 19的小鼠进行仔细的组织病理学分析， 找出围产期死亡的原因这将包括 肺、心脏和骨髓中细胞类型的鉴定 细胞，其中ADAM19是高度表达，并在体内研究，以评估 这些细胞和组织中的ADAM19的功能。 2)解决功能冗余和补偿的可能性， ADAM19和其他ADAM在发育过程中通过产生缺乏其他 除了ADAM 19之外，还有其他相关的ADAM。 3)在缺乏ADAM 19或ADAM19的细胞中发现胞外域脱落的潜在选择 使用有偏和无偏的方法将其他ADAM与野生型细胞进行比较。 有偏见的方法将集中在蛋白质，已知是脱落， 有助于缺乏ADAM 19的小鼠的表型，如EGFR-配体、TNF-α、TNF-α和TNF-α。 和TNF受体家族成员。无偏的方法将比较蛋白质 分泌到野生型和ADAM19敲除细胞的上清液中。这 这项研究将提供令人兴奋的见解ADAM19的功能， 发育和成年小鼠，并将揭示缺陷的机制， 导致围产期死亡。我们的研究结果很可能 对人类疾病中ADAM 19功能的影响，并可能揭示新的 在由疾病引起的疾病中进行治疗干预的目标 错误调节或有缺陷的ADAMI9或其他相关的ADAMs。

DESCRIPTION (provided by applicant): Metalloprotease-disintegrins (ADAMs) are a family of membrane anchored glycoproteins that have key roles in fertilization, neurogenesis, and in activation of EGF receptor ligands during development. The main goal of this proposal is to uncover the mechanism underlying the perinatal lethal phenotype of mice lacking the widely expressed ADAM 19, with an emphasis on evaluating the predicted role of ADAM19 in protein ectodomain shedding. Protein ectodomain shedding leads to the release of a variety of structurally and functionally diverse proteins from the plasma membrane, and has recently emerged as a critical post-translational mechanism of regulating the function of the released proteins. Ectodomain shedding is essential for proper signaling via EGF receptor-ligands, Notch-mediated lateral inhibition, limiting TNF receptor mediated inflammatory reactions, and regulating several aspects of axonal guidance. Because the first identified sheddase, the TNFalpha convertase, is an ADAM, and because lIMP inhibition studies rule out currently known matrix metalloproteases (MMPs) as sheddases for most proteins, ADAMs have emerged as the best candidate ectodomain sheddases. Our working hypothesis is that the phenotype of mice lacking ADAM19 is due, at least in part, to a defect in ectodomain shedding of one or more proteins. The specific aims of this project are: 1) Perform a careful histopathological analysis of mice lacking ADAM19 in order to uncover the cause of the observed perinatal lethality. This will include an identification of the cell types in the lung, heart, and in bone marrow derived cells where ADAM19 is highly expressed, and in vivo studies to evaluate the function of ADAM19 in these cells and tissues. 2) Address the possibility of functional redundancy and compensation between ADAM 19 and other ADAMs during development by generating mice lacking other related ADAMs in addition to ADAM19. 3) Uncover potential aelects in ectodomain shedding in cells lacking ADAM 19 or other ADAMs compared to wildtype cells using a biased and unbiased approach The biased approach will focus on proteins that are known to be shed and could contribute to the phenotype of mice lacking ADAM 19, such as EGFR-ligands, TNF- and TNF-receptor family members. The unbiased approach will compare proteins secreted into the supernatant of wildtype and ADAM 19 knockout cells. This study will provide exciting insights into the function of ADAM19 during development and in adult mice, and will uncover the mechanism by which a defect in ADAM 19 leads to perinatal lethality. Our results are likely to have implications for the function of ADAM19 in human disease, and may uncover novel targets for therapeutic interventions in diseases that are caused by a misregulated or defective ADAMI9 or by other related ADAMs.