SUBEPITHELIAL ANTIGENS

上皮下抗原

• 批准号: 2542941
• 负责人: ROBERT E BURGESON
• 金额: $ 35.72万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-23 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2542941
• 关键词: artificial skin; basement membrane; biological models; cell adhesion; cell adhesion molecules; epidermolysis bullosa; epithelium; human tissue; intercellular connection; keratinocyte; laboratory mouse; laboratory rabbit; laminin; protein sequence; protein structure function; skin

The anchoring complex is an ultrastructural feature of the dermal-epidermal basement membrane that includes the hemidesmosomes, anchoring filaments and anchoring fibrils. These structures are believed to work as a unit to stabilize epithelial attachment to the underlying basement membrane. Failure of components of the anchoring complex results in spontaneous blistering characteristic of the inherited bullous diseases. We have recently identified two laminin variants that are components of the anchoring filaments. We have termed these kalinin and K-laminin. We know that kalinin is absent from the basement membrane zone of patients with lethal epidermolysis bullosa. Kalinin is a cell adhesion macromolecule specific for certain types of epithelial cells including keratinocytes. For these cells, kalinin substitutes for laminin as the favored growth substrate. Monoclonal antibodies made to kalinin cause cell detachment and de-epithelialization of whole skin in vitro. This application is focused upon continued characterization of the structure and function of kalinin and K-laminin. We propose to isolate kalinin and K-laminin for protein sequence determinations and for preparing protein fragments; to complete the deduced amino acid sequences of these chains and to express selected subdomains in vitro; and to generate domain specific monoclonal antibodies. These reagents will be used to evaluate the binding function of kalinin and K-laminin subdomains. We will interfere with these functions in an in vitro skin equivalent model. These probes will also be used to evaluate mutations in the kalinin and K-laminin chains relative to the pathogenesis of junctional epidermolysis bullosa.

锚定综合体是 皮肤表皮基底膜，包括半体体， 锚固细丝和锚定纤维。 相信这些结构 作为一个单位工作以稳定上皮附件 地下室膜。 锚定复合物的组件故障 导致遗传大胆的自发起泡特征 疾病。 我们最近确定了两个层粘连蛋白变体 锚固丝的组成部分。 我们称这些kalinin和 k-甲胺素。 我们知道基底膜不存在Kalinin 致死性表皮溶解的患者区域。 kalinin是一个细胞 特异于某些类型上皮细胞的粘附大分子 包括角质形成细胞。 对于这些细胞，kalinin替代 层粘连蛋白是受青睐的生长底物。 单克隆抗体 kalinin在整个皮肤中引起细胞脱离和去皮上的上皮化 体外。 该应用的重点是持续表征的 kalinin和k层蛋白的结构和功能。 我们建议 用于蛋白质序列确定的分离酸蓝氨酸蛋白和K-氯胺酮 制备蛋白质碎片；完成推导的氨基酸序列 在这些链中，并在体外表达选定的子域；然后 产生特定的域单克隆抗体。 这些试剂将是 用于评估kalinin和k-果岭蛋白的结合功能 subdomains. 我们将在体外皮肤中干扰这些功能 等效模型。 这些探针也将用于评估突变 在kalinin和k层蛋白链中相对于的发病机理 连接表皮溶解Bullosa。