BIOLOGY OF HYALURONIDASES

透明质酸酶是一类具有中性和酸性活性的酶， 在整个动物王国，关键的基本过程， 细胞运动性、胚胎发育、伤口愈合、血管生成和 肿瘤发生 尽管这些酶显然是无效的，但它们并不是 由于缺乏快速、灵敏的检测， 这 随着我们开发ELISA样测定法，这个问题已经克服 足够敏感，以便可以进行动力学，并且活性 小样本可以测量。 在一项器官调查中， 在肝、肺、肾和脾中发现活性。 肝酶 以及血清酶现在已经被纯化到同质， 不同的蛋白质。 Hybronidases似乎是一个广泛的家族， 酶蛋白质。 我们建议分离这个家族的其他酶， 并利用协调的生化免疫和分子生物学 方法，检查每个生物学，识别前体形式， 在培养细胞和组织中建立精确的组织定位 路段 生物素化HA结合蛋白探针还将允许 同时显示基质。 我们预测， 系统，如脾酶的例子，是主要的分解代谢途径 对于HA。 肝脏和肾脏负责额外的 分解代谢途径，游离HA在循环中可能是一个 分级取决于HA聚合物尺寸。 成纤维细胞中的透明质酸酶 目前，通过直接测定和利用Western blot检测， 针对纯化的肝酶的多克隆抗体。 检测到这种中性酶活性，以前在 正常哺乳动物成纤维细胞，通过提取 存在高盐，这似乎会分解酶抑制剂 复杂. 我们将使用适当的cDNA文库进行测序 透明质酸酶。 共有序列，以及模块化结构域 将在这个新的酶家族中鉴定出模式。 为什么会有这样 脊椎动物组织中的透明质酸酶的广泛库？ 为什么 HA底物本身以这样多种状态存在：游离于 循环、细胞结合、完全水合或与光谱相关 HA结合蛋白。 为了回答这些问题，我们将 试图将酶动力学和透明质酸酶模式相关联 各种组织中的蛋白质结构与分子状态的位置 伴随的HA基质和组织特异性的要求 功能协调发展的

Hyaluronidases are a group of neutral- and acid- active enzymes found throughout the animal kingdom, critical to such fundamental process as cell motility, embryological development, wound healing, angiogenesis and tumorigenesis. Despite their obvious impotence, these enzymes are not well understood because of the lack of a rapid, sensitive assay. This problem has now been overcome with our development of an ELISA-like assay sufficiently sensitive so that kinetics can be performed, and activity in small samples can be measured. In an organ survey, high levels of activity were found in liver, lung, kidney and spleen. The liver enzyme as well as the serum enzyme have now been purified to homogeneity an dare indeed different proteins. Hyaluronidases appear to be a broad family of enzyme proteins. We propose to isolate other enzymes of this family, and using a coordinated biochemical, immunological, and molecular biology approach, examine the biology of each, identify precursors forms, and establish precise histolocalization, both in cultured cells and in tissue sections. The biotinylated HA-binding protein probe will also permit simultaneous visualization of substrate. We predict that the lymphatic system, as exemplified by the spleen enzyme, is the major catabolic route for HA. The liver and the kidneys are responsible for additional catabolic pathways, of free HA in the circulation in what may be a hierarchy dependent on HA polymer size. The hyaluronidase in fibroblasts has also been detected now, by direct assay and by Western blot utilizing the polyclonal antibodies directed against the purified liver enzyme. detection of this neutral enzyme activity, not observed previously in normal mammalian fibroblasts, was facilitated by extracting in the presence of high salt, which appears to disaggregate an enzyme-inhibitor complex. We will use appropriate cDNA libraries to sequence hyaluronidase enzymes. Consensus sequences, as well as modular domain patterns will be identified in this new enzyme family. Why is there such a wide repertoir of hyaluronidase in vertebrate tissue? And why does the HA substrate itself exist in such a wide variety of states: free in the circulation, cell-bound, fully hydrated or in association with a spectrum of HA-binding proteins. Toward an answer to such questions, we will attempt to correlate enzyme kinetics and patterns of hyaluronidase protein structure in various tissues with the molecular state an location of the attendant HA substrate and the requirements for tissue-specific functions.