KERATINOCYTE NEUTRAL SECRETORY PROTEASES AND INHIBITORS

角质细胞中性分泌蛋白酶和抑制剂

• 批准号: 3219758
• 负责人: HENNING BIRKEDAL-HANSEN
• 金额: $ 12.99万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 1986-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219758
• 关键词: basement membrane; cell differentiation; collagenase; cyclic nucleoside monophosphate; glucocorticoids; heparin; inflammation; keratinization; peptidases; plasminogen; plasminogen activator; protease inhibitor; proteoglycan

A study is proprosed of the function and regulation of neutral secretory proteases in keratinocyte-mediated interstitial collagen and basement-membrane degradation. (1) The ability of live keratinocytes to degrade interstitial type I and III collagen fibrils in culture will be measured and the specific and coordinate role of the component proteases of the collagenolytic complex: a 65K collagenase, a 110K gelatinase, two (70K and 48K) plasminogen activators and exogenous plasminogen will be analyzed by immune-inhibition. A search will be made for additional components of the collagenolytic complex such as specific activators of procollagenase and plasminogen proactivators and the regulatory role of plasma and endogenous secretory protease inhibitors, including epidermal inhibitors of the fibrinolytic and collagenolytic systems, will be analyzed. (2) The ability of keratinoyctes to degrade intact basement membranes will be investigated. The component proteases of the type IV collagen, laminin, fibronectin and heparan sulfate proteoglycan degrading systems will be identified and the specific function of the type IV-collagen-degrading protease system will be analyzed by specific immune inhibition. (3) The expression of proteolytic competence as a function of basal cell activation will be investigated. (a) The inverse relationship between secretion of specific proteases and terminal differentiation induced by anchorage deprivation in vitro will be established and the distribution and position of secretor cells in surface-attached primary explants and in clonal colonies will be correlated with that of differentiation markers. (b) The synthesis of proteases by activated basal cells induced by excision wounding in vivo will be studied by immunofluorescence and correlated with migration, confluence and stratification during the healing process. (c) The effect on interstitial collagen and basement membrane degradation of agents which modulate expression of activated and differentiated cellular states such as retinoids, glucocorticoids, inflammatory agents and cyclic nucleotides will be analyzed.

本文对中性分泌型胶原的功能和调节进行了研究。 角质形成细胞介导的间质胶原蛋白中的蛋白酶， 基底膜降解 (1)活的角质形成细胞能够 在培养物中降解间质I型和III型胶原原纤维将是 测量和组分蛋白酶的特定和协调作用， 胶原溶解复合物：一种65K胶原酶，一种110K明胶酶，两种（70K 和48K）纤溶酶原激活剂和外源性纤溶酶原进行分析 免疫抑制。 将搜索以下内容的其他组成部分： 胶原溶解复合物如前胶原酶的特异性激活剂 和纤溶酶原激活剂的调节作用， 内源性分泌蛋白酶抑制剂，包括表皮抑制剂， 纤维蛋白溶解和胶原溶解系统。 (2)的 角化菌降解完整基底膜的能力将是 研究了 IV型胶原蛋白的组分蛋白酶，层粘连蛋白， 纤维连接蛋白和硫酸乙酰肝素蛋白聚糖降解系统将是 鉴定和IV型胶原降解的具体功能， 蛋白酶系统将通过特异性免疫抑制进行分析。 (3)的 作为基底细胞活化的函数的蛋白水解能力的表达 将进行调查。 (a)分泌的负相关性， 特异性蛋白酶与锚定诱导的终末分化 体外剥夺将建立和分布和位置 分泌细胞在表面附着的原代外植体和克隆 集落将与分化标志物的集落相关联。 (b)的 通过切除诱导的活化基底细胞合成蛋白酶 将通过免疫荧光研究体内创伤，并与 愈合过程中的迁移、汇合和分层。 （c）第（1）款 对间质胶原和基底膜降解的影响 调节活化和分化的细胞因子表达的试剂 状态，如类维生素A、糖皮质激素、炎症剂和环状 将分析核苷酸。