I型IL-1受体的结构与其转运功能相关的研究  

The relationship between the IL-1 receptor structure and its trafficking function

作　　者：黄文燕[1] 罗兆明[1] David Wyllie Eva E.Qwarnstrom 

机构地区：[1]广州医学院人体解剖学教研室,广东广州510182 [2]Molecular Cell Biology,Division of Molecular and Genetic Medicine,The University of Sheffield,UK

出　　处：《中国病理生理杂志》2001年第2期112-115,共4页Chinese Journal of Pathophysiology

摘　　要：目的 :为进一步了解IL - 1受体的结构与IL - 1受体在细胞内表达后的转运的关系 ,以及该结构改变对IL - 1受体功能的影响。方法 :将重组型、突变型和仅含细胞外段的I型IL - 1受体 (IL - 1RI)的质粒构建入含有绿色荧光蛋白的表达载体中 ,用磷酸钙沉淀法转染入成纤维细胞中 ,通过激光扫描共聚焦显微镜 ,观察其表达的融合蛋白在单个活细胞中的定位 ,以及与IL - 1结合后的运动变化。结果 :重组型IL - 1RI的融合蛋白表达后主要位于细胞膜和细胞伸出的突起与细胞基质相接触的焦点附着斑处 ,加入IL - 1后 ,逐渐从细胞膜上移至细胞质内 ,最后聚集在细胞核上。突变型IL - 1RI和细胞外段IL - 1RI的融合蛋白表达后散布于细胞质内 ,对IL - 1的剌激不产生反应。结论 :纤维粘连蛋白可增加IL - 1RI的表达转染率。IL - 1RI表达后的转运过程与IL - 1RI细胞内段结构域的完整和精确有关 ,尤其需要C -末端第 5 14位色氨酸的参与。AIM:To analyze the relationship between structure and trafficking function of IL-1 receptor.METHODS: Wild type and mutant (W514A) IL-1 receptor cDNA containing mouse extracellular domain and human intracellular domain, IL-1 receptor cDNA only containing mouse extracellular domain, were inserted into pEGFP-N2 C-terminal protein fusion vector, respectively. All constructs were transfected respectively into human fibroblasts using calcium phosphate precipitation method. The location of expressing fusion protein at single cell level and translocation after binding with IL-1 were determined under confocal microscopy. RESULTS: The wild type IL-1 receptor located in cell membrane, extended processes and accumulated in areas suggestive of sites of focal adhesions. Further, the fusion proteins moved into cytoplasm and finally concentrate in nuclear after IL-1 stimulation. In contrast, mutant and extracellular domain IL-1 receptors did not translocate to sites of cell adhesion and were unaffected by IL-1 stimulation. CONCLUSION: Fibronectin can increase the expression of IL-1 receptor. The trafficking of IL-1 receptor is dependant on structurally intact, specific structure (tryptophan at 514) of C-terminal end of the cytoplasmic tail of the receptor is necessary.

关 键 词：受体 白细胞介素l 突变 生物转运 

分 类 号：Q26[生物学—细胞生物学]