Nuclear Localization of Heparan Sulfate Proteoglycans

硫酸乙酰肝素蛋白多糖的核定位

• 批准号: 6724814
• 负责人: MATTHEW A NUGENT
• 金额: $ 16.15万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6724814
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; cell nucleus; corneal stroma; extracellular matrix proteins; eye injury; fibroblasts; heparan sulfate; image processing; intracellular transport; ion exchange chromatography; organ culture; protein kinase C; protein transport; proteoglycan

DESCRIPTION: (Applicant's Abstract) Stromal injury results invariably in stromal scarring and in many instances impaired vision. Many in vivo and in vitro models have been used to study corneal wound healing. Importantly, evidence from these studies suggests that there are certain common biochemical events that occur in response to injury. Foremost amongst these are changes in the composition and organization of the stromal extracellular matrix. In addition, the presence of the glycosaminoglycan, heparan sulfate, within the corneal stroma has been noted in a number of injury models. The corneal stroma. is normally devoid of heparan sulfate, suggesting that its presence at sites of injury represents an integral component of the cellular response to injury. We have recently noted that the intracellular localization of heparan sulfate can be regulated by the state of the extracellular matrix. We found that a large fraction of the heparan sulfate proteoglycan (HSPG) expressed by corneal fibroblasts in culture is localized to the nucleus when cells were grown on a fibronectin containing matrix. Thus, the overall goals of this proposal are to identify the mechanisms and consequences of HSPG nuclear localization in stromal fibroblasts. We hypothesize that the nuclear translocation of HSPG is regulated by interactions between extracellular proteins and heparan sulfate chains. We propose that these interactions stimulate intracellular signaling events involving protein kinase C family members. We will evaluate possible consequences of nuclear HSPG including the likelihood that HSPG can be used as a macromolecular shuttle for extracellular proteins to access the nucleus. The specific aims of this proposal are: 1. Define the mechanisms underlying HSPG nuclear translocation, and 2. Identify potential functional consequences of nuclear HSPG. These studies will have wide ranging implications throughout eye research and could reveal a previously undetected generalized mechanism for nuclear-extracellular communication. Information from these studies could provide the foundation for the development of a new generation of therapies aimed at facilitating corneal repair without scarring.

描述：（申请人的摘要）基质损伤结果总是在 基质疤痕，在许多情况下，视力受损。许多体内和 体外模型已用于研究角膜伤口愈合。重要的是， 这些研究的证据表明，存在某些常见的生化 响应伤害的事件。其中最重要的是变化 基质外基质的组成和组织。在 此外，在糖胺聚糖乙酰乙酰胺硫酸盐的存在中存在 在许多损伤模型中已注意到角膜基质。角膜基质。 通常没有硫酸乙酰肝素，表明其存在于 损伤代表了细胞对损伤的反应的组成部分。我们 最近指出，硫酸乙酰肝素的细胞内定位可以 受细胞外基质状态的调节。我们发现一个大 角膜表达的硫酸乙酰肝素蛋白聚糖（HSPG）的比例 当细胞生长在A上时，培养中的成纤维细胞位于核中 纤连蛋白含有基质。因此，该提议的总体目标是 确定HSPG核定位的机制和后果 基质成纤维细胞。我们假设HSPG的核易位是 受细胞外蛋白与硫酸乙酰肝素之间的相互作用的调节 链。我们建议这些相互作用刺激细胞内信号传导 涉及蛋白激酶C家族成员的事件。我们将评估可能 核HSPG的后果，包括HSPG可以用作的可能性 细胞外蛋白的大分子班车进入细胞核。这 该提案的具体目的是：1。定义HSPG基础机制 核转运和2。确定潜在的功能后果 核HSPG。这些研究将在整个眼中具有广泛的影响 研究，可以揭示先前未发现的通用机制 核 - 细胞交流。这些研究的信息可能 为开发新一代疗法提供基础 旨在促进角膜维修而无需疤痕。