C1 FUNCTION ANALYZED WITH NATURALLY-OCCURRING INHIBITORS

使用天然抑制剂分析 C1 功能

• 批准号: 2139208
• 负责人: ANNE NICHOLSON-WELLER
• 金额: $ 19.78万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2139208
• 关键词: SDS polyacrylamide gel electrophoresis; blocking antibody; carbohydrate sequence; complement; complement inhibitors; complement pathway; gene expression; glycosidases; heparan sulfate; high performance liquid chromatography; human tissue; laboratory mouse; laboratory rabbit; mass spectrometry; monoclonal antibody; posttranslational modifications; protein biosynthesis; protein purification; protein sequence; protein structure function

When immune complexes bind the C1q subcomponent of C1, the bound C1q has two important activities: 1) providing a binding site for C1r2s2, the catalytic unit of C1, which activates the rest of the complement cascade; and 2) stimulating cells bearing the C1q receptor. The overall objectives of this proposal are to identify and characterize the natural inhibitors of C1, which are responsible for the specific and restricted functional activity of C1 in vivo. In particular, we are focusing on two inhibitors, Factor J and heparan sulfate, which are potentially able to control the recycling of bound C1q as an activator of Clr2s2, and as a stimulus to cells bearing C1q receptors. Factor J was initially identified and isolated from human urine and serum in the Principal Investigator's laboratory. It is a heavily glycosylated, basic protein, comprised of 200,000, 18,000, and 5,900 subunits, which is expressed on some circulating cells. Factor J binds to C1q and is a potent non-competitive inhibitor of the assembly of macromolecular C1 from the C1q and C1r2s2 subcomponents. It is structurally and functionally distinct from other known inhibitors of C1. The protein sequence of Factor J will be determined by Edman degradation or mass spectrometry using proteolytic fragments of deglycosylated protein. By producing high titer polyclonal and monoclonal antibodies it will be possible to define the functional and antigenic domains of the protein, characterize the expression of Factor J antigen in various tissues, and perform biosynthetic studies.

当免疫复合物结合C1的C1 q亚组分时，结合的C1 q具有 两个重要的活性：1）提供C1 r2 s2的结合位点， C1的催化单元，其激活补体级联的其余部分; 和2）刺激带有C1 q受体的细胞。 总体目标 这项建议的目的是确定和表征天然抑制剂， C1，负责特定和受限功能 体内C1活性。 我们特别关注两种抑制剂， 因子J和硫酸乙酰肝素，这是潜在的能够控制 结合的C1 q的再循环作为Clr 2s 2的激活剂，并作为刺激， 携带C1 q受体的细胞。 J因子最初是从人的尿液和血清中鉴定和分离出来的 在主要研究者的实验室里。 它是一种高度糖基化的， 碱性蛋白，由200，000、18，000和5，900个亚基组成， 在一些循环细胞上表达。 因子J与C1 q结合，是一种有效的 从C1 q组装大分子C1的非竞争性抑制剂 和C1 r2 s2子组分。 它在结构和功能上都是不同的 其他已知的C1抑制剂。 因子J的蛋白质序列将 通过Edman降解或使用蛋白水解的质谱法测定 去糖基化蛋白的片段。 通过生产高滴度多克隆 和单克隆抗体，将有可能定义功能性和 蛋白质的抗原结构域，表征因子J的表达 抗原在各种组织中，并进行生物合成研究。