GLYCOPROTEINS IN DESQUAMATION IN SKIN AND ORAL MUCOSA

皮肤和口腔粘膜脱屑中的糖蛋白

• 批准号: 2130049
• 负责人: MIRIAM M BRYSK
• 金额: $ 15.94万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1996-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2130049
• 关键词: cell differentiation; enzyme structure; gene expression; genetic transcription; glycoproteins; human tissue; interferon gamma; keratinocyte; laboratory rabbit; lectin; molecular biology; northern blottings; nucleic acid sequence; oral mucosa; serine proteinases; skin; tissue /cell culture

The research in this proposal continues our studies of the role of glycoproteins in epidermal desquamation, with emphasis on three related serine proteinases that we have isolated from human epidermis (and also find in the more differentiated cells of the oral cavity). Predesquamin (600 kD), immunolocalized to the upper viable layers and the lower stratum corneum, has both thrombin and tryptase activities. It is the precursor of a 400 kD molecule with thrombin activity and of a 40 kD glycoprotein (desquamin) with tryptase activity. Desquamin, immunolocalized to the stratum corneum only, is also a lectin. We showed in cell aggregation experiments that desquamin affects cohesion and desquamation, and we believe that all three molecules are crucial to the degradative events of terminal differentiation. They are not ordinarily expressed in vitro, but we have developed a culture system in which interferon-gamma induces their expression. We will determine to what extent our three enzymes can degrade various structural components of the stratum corneum and whether they act in successive stages. We will elucidate the processing of predesquamin to the other two (and any intermediates) from structural and functional correlations. In particular, we will completely sequence desquamin biochemically, partially sequence the other two, and seek homology with known serine proteinases. Polyclonal antibodies to selected peptides will be used to immunoprecipitate intermediates and for immunolocalization in different cell populations. Given the amino acid sequences of degraded peptides, we will develop oligonucleotide probes to clone for the gene. We will study gene expression, in normal and diseased tissue, and its modulation by agents known to affect proliferation and differentiation, in vitro (with our interferon-gamma system) and in vivo.

本提案中的研究继续我们对以下角色的研究： 表皮脱落中的糖蛋白，重点是三个相关的 我们从人类表皮中分离出丝氨酸蛋白酶（以及 发现于口腔分化程度较高的细胞中）。 前去角蛋白 (600 kD)，免疫定位于上层活细胞层和下层细胞 角质层，具有凝血酶和类胰蛋白酶活性。 它的前身是 具有凝血酶活性的 400 kD 分子和 40 kD 糖蛋白 （脱屑素）具有类胰蛋白酶活性。 Desquamin，免疫定位于 角质层而已，也是一种凝集素。 我们在细胞聚集中展示 实验表明脱屑素影响内聚力和脱屑，我们 相信所有这三个分子对于降解事件都至关重要 终端分化。 它们通常不在体外表达，但 我们开发了一种培养系统，其中干扰素γ诱导它们 表达。 我们将确定我们的三种酶可以降解到什么程度 角质层的各种结构成分及其是否起作用 在连续的阶段中。 我们将阐明前脱屑素的加工过程 其他两个（以及任何中间体）来自结构和功能 相关性。 特别是，我们将彻底测序脱皮蛋白 生化上，对另外两个进行部分测序，并寻求与 已知的丝氨酸蛋白酶。 针对所选肽的多克隆抗体将 用于免疫沉淀中间体和免疫定位 不同的细胞群。 给定降解的氨基酸序列 肽，我们将开发寡核苷酸探针来克隆该基因。 我们 将研究正常组织和患病组织中的基因表达及其 已知影响增殖和分化的物质的调节， 体外（使用我们的干扰素-γ系统）和体内。