INTERACTIONS OF GELATINASES WITH SERUM/MEMBRANE PROTEIN

肿瘤细胞的转移性扩散通常是 癌症患者。对分子的更完整的理解 转移机制是找到治愈任何形式的肿瘤的关键。 恶毒。明胶酶A和B直接参与肿瘤转移。这个 酶在基底物的溶解中起着重要作用。 在入侵的最初阶段，膜等是至关重要的 渗出。 这项提议的广泛的长期目标有两个。首先， 描述明胶酶之间结合相互作用的意义 和血清糖蛋白，积雪草素和胎球蛋白，和 另一方面，明胶酶及其可能的膜受体。 其次，为了表明膜结合明胶酶是否具有其他功能 管家的作用，除了溶解基底膜。 具体目的是：a)阐明两者中的结合结构域 去唾液酸叶绿素和明胶酶，b)评价可溶性和 固定化积雪草素/胎球蛋白及其他相关蛋白 激活前明胶酶，c)识别细胞膜受体 对于明胶酶和评估配体-受体的相互作用，d)到 明胶酶与31 kDa的细胞表面相互作用分析 人半乳糖苷结合蛋白(人半乳凝素-3)。放射性标记 冷的前明胶酶将与胎球蛋白/积雪草素和 被监测的酵母原的结合、切割和激活。血浆 膜受体也将被分离、纯化和配体-受体 相互作用已分析。这类研究将提供有关 明胶酶在细胞表面和细胞内的活性 细胞内空间。

The metastatic spread of tumor cells is usually the cause of death in cancer patients. A more complete understanding of the molecular mechanisms of metastasis is pivotal to finding a cure for any form of malignancy. Gelatinases A and B are directly involved in metastasis. The enzymes play a significant role in the dissolution of the basement membrane and so are critical in the initial stages of invasion and extravasation. The broad long-term objectives of this proposal is twofold. Firstly, to delineate the significance of the binding interactions between gelatinases and the serum glycoproteins, asialofetuin and fetuin on the one hand, and gelatinases and their putative membrane receptors on the other hand. Secondly, to show whether or not membrane bound gelatinases perform other housekeeping roles, apart from the dissolution of the basement membrane. The specific aims are: a) To elucidate the binding domains in both asialofetuin and gelatinases, b) To evaluate the ability of soluble and immobilized asialofetuin/fetuin and other related proteins in the activation of pro-gelatinases, c) To identify the cell membrane receptors for gelatinases and evaluate the ligand-receptor interactions, d) To analyze the cell surface interactions between gelatinases and the 31 kDa human galactoside binding protein (human galectin-3). Radioactive labeled and cold pro-gelatinases will be incubated with fetuin/asialofetuin and binding, cleavage, and activation of the zymogens monitored. The plasma membrane receptors will also be isolated, purified and the ligand-receptor interactions analyzed. Such studies will provide vital information on the activity of gelatinases on the cell surface as well as in the intracellular spaces.