STRUCTURE AND FUNCTION OF THE INTEGRIN ALPHA1 BETA1

整合素 ALPHA1 BETA1 的结构和功能

• 批准号: 3303765
• 负责人: EUGENE E. MARCANTONIO
• 金额: $ 18.66万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3303765
• 关键词: antibody formation; basement membrane; cell cell interaction; cell differentiation; cell migration; chimeric proteins; collagen; embryo /fetus cell /tissue; extracellular matrix; human tissue; in situ hybridization; integrins; laboratory mouse; laboratory rabbit; laminin; molecular cloning; nucleic acid probes; protein structure function; receptor binding; site directed mutagenesis; tissue /cell culture

The main objective of the proposed research plan is to investigate the mechanisms by which cell control their interactions with extracellular matrix and basement membranes during migration and differentiation in the normal and disease state. The integrin receptor heterodimer alpha1-beta1, a laminin/collagen receptor, has been chosen as a model system for this study because of several unique features. This molecular has a limited distribution in vivo, being present on activated lymphocytes, neuronal cells and in the mesangium of the kidney in adult tissues studied to date. This distribution implies a selective and specific function which is not fulfilled by other integrin collagen/laminin receptors (i.e., alpha2,3,and 6). This integrin can be induced on the surface of lymphocytes after activation in vitro, on PC12 cells in vitro, and in vivo on the synovial lymphocytes of patients with rheumatoid arthritis. Thus, this molecule probably is tightly regulated in vivo, and enables us to study the control of integrin function. cDNA clones for the human integrin alpha1 subunit will be used to produce fusion proteins and synthetic peptides for the generation of antibodies. The role of alpha1 in providing positional information in cell migrations crucial to normal development will be investigated by determining the temporal and spatial pattern of expression in mammalian embryos suing these probes. In particular, the presence of this integrin on migrating cells will allow us to determine if there are changes in expression levels from early in development, when the cells are migrating, to late, when they have reached their destination. Lastly, this integrin has shown different ligand specificities on varying cell types, binding laminin and collagen on some, while binding only collagen on others, a property which permits investigation of the factors both intrinsic and extrinsic to alpha1 which affect ligand specificity in cell migrations in vivo. These factors will be studied using intact, truncated and mutant forms of the molecule, the latter two generated by site-directed mutagenesis and heterologous eukaryotic expression systems. This research will enhance our knowledge of the mechanism of cell migration in both normal states such as development, and abnormal states such as tumor cell metastasis, and chronic inflammatory diseases; with the long term goal of determining the role of alpha1 and other integrins in the targeting of cells in disease states.

拟议的研究计划的主要目的是调查 细胞控制其与细胞外相互作用的机制 迁移和分化过程中的基质和基底膜 正常和疾病状态。 整联蛋白受体异二聚体alpha1-beta1， 层粘连蛋白/胶原蛋白受体已被选为模型系统 由于几个独特的特征而学习。 该分子有限 体内分布，存在于活化的淋巴细胞上，神经元 迄今为止研究的成人组织中的细胞和肾脏中的肾脏中。 此分布意味着一个选择性和特定功能不是 由其他整合素胶原蛋白/层粘连蛋白受体（即α2,3和 6）。 该整联蛋白可以在淋巴细胞表面诱导 体外激活，体外PC12细胞和滑膜上的体内激活 类风湿关节炎患者的淋巴细胞。 因此，这个分子 可能在体内受到严格调节，使我们能够研究控制 整联蛋白功能。 人整合素alpha1亚基的cDNA克隆 将用于生产融合蛋白和合成肽 产生抗体。 alpha1在提供位置的作用 细胞迁移中对正常发育至关重要的信息将是 通过确定表达的时间和空间模式来研究 在提起这些探针的哺乳动物胚胎中。 特别是存在 迁移细胞上的整合素将使我们能够确定是否存在 从发育初期开始的表达水平的变化，当细胞为 当他们到达目的地时，迁移到迟到。 最后，这个 整合素在不同的细胞类型上显示出不同的配体特异性， 结合层粘连蛋白和胶原蛋白，同时仅结合胶原蛋白 其他的，允许对两者的因素进行调查的属性 影响细胞中配体特异性的固有和外在的α1 体内迁移。 这些因素将使用完整，截断 分子的突变形式，后两个由位置定向产生 诱变和异源真核表达系统。 这项研究 将增强我们对两者中细胞迁移机制的了解 正常状态，例如发育和异常状态，例如肿瘤细胞 转移和慢性炎症性疾病；长期目标的 确定α1和其他整合素在靶向的作用 疾病状态中的细胞。