PHOSPHORYLATION OF CALPAINS

钙蛋白酶的磷酸化

• 批准号: 8171284
• 负责人: DARREL E GOLL
• 金额: $ 0.08万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171284
• 关键词: Alzheimer' s Disease; Area; Blood Vessels; Calpain; Cataract; Cell physiology; Cells; Computer Retrieval of Information on Scientific Projects Database; Crystallins; Event; Funding; Genes; Goals; Grant; Institution; Learning; Life; Muscular Dystrophies; Myocardial Infarction; Normal Cell; Pathology; Peptide Hydrolases; Phosphorylation; Regulation; Research; Research Personnel; Research Project Grants; Resources; Site; Source; Spinal cord injury; Stroke; Tissues; United States National Institutes of Health; in vitro Assay; m-calpain; muscle form

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long-term goal of this research is to learn how activity of the ubiquitous calpains, ?- and m-calpain, is regulated in living cells. The calpains are Ca2+-dependent proteases found in every vertebrate cell. Disruption of the gene encoding the 28-kDa subunit common to ?- and m-calpain is embryonically lethal. Inappropriate calpain degradation is implicated in many tissue pathologies ranging from loss of muscle mass in the muscular dystrophies, to crystallin degradation and cataract formation, to Alzheimer?s disease, to tissue damage in ischemic areas near a blocked blood vessel (stroke or myocardial infarction), to traumatic spinal cord injury, etc. Calpain activity in these pathologies is triggered by elevated, intracellular [Ca2+]; however, Ca2+ requirement of the calpains in in vitro assays is 3-50 ?M (?-calpain) or 300-500 ?M (m-calpain), much higher than intracellular free [Ca2+] is, even near ischemic areas. Cells, therefore, have a mechanism to reduce the [Ca2+] required for calpain activity. This mechanism evidently is altered in a way that activates the calpains during ischemic events. The same mechanism must regulate calpain activity during normal cell function. We have found that both ?- and m-calpain are phosphorylated at multiple sites. The objective of our current research project is to identify the phosphorylated sites on both ?- and m-calpain and determine their effect on regulation of calpain activity.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 这项研究的长期目标是了解普遍存在的钙蛋白酶、β-和 m-钙蛋白酶的活性如何在活细胞中受到调节。钙蛋白酶是每个脊椎动物细胞中都存在的 Ca2+ 依赖性蛋白酶。编码α-和m-钙蛋白酶共有的28-kDa亚基的基因的破坏是胚胎致死的。不适当的钙蛋白酶降解与许多组织病理有关，包括肌营养不良症中的肌肉质量损失、晶状体蛋白降解和白内障形成、阿尔茨海默氏病、血管阻塞附近缺血区域的组织损伤（中风或心肌梗塞）、创伤性脊髓损伤等。这些病理中的钙蛋白酶活性是 由细胞内 [Ca2+] 升高触发；然而，体外测定中钙蛋白酶的Ca2+需求为3-50μM（β-钙蛋白酶）或300-500μM（m-钙蛋白酶），远高于细胞内游离[Ca2+]，甚至在缺血区域附近也是如此。因此，细胞具有减少钙蛋白酶活性所需的[Ca2+]的机制。这种机制显然以在缺血事件期间激活钙蛋白酶的方式发生改变。在正常细胞功能期间，相同的机制必须调节钙蛋白酶活性。我们发现α-和m-钙蛋白酶在多个位点被磷酸化。我们当前研究项目的目标是确定 β- 和 m-钙蛋白酶上的磷酸化位点，并确定它们对钙蛋白酶活性调节的影响。