抗体交联作用促进Nephrin簇集于细胞膜脂筏微区  

Nephrin Clustered in Lipid Raft-associated Microdomain on the Cell Membrane by Antibody Induced Cross-linking

作　　者：秦晓松[1] 刘勇[1] 佟威威[1] 岳丹[1] 刘建华[1] 刘岩[1] 

机构地区：[1]中国医科大学附属盛京医院检验科,沈阳110004

出　　处：《中国细胞生物学学报》2010年第2期241-245,共5页Chinese Journal of Cell Biology

基　　金：辽宁省自然科学基金(No.20082095);辽宁省高校科研计划项目资助~~

摘　　要：为研究nephrin在细胞膜上的表达特点,构建nephrin和podocin的表达质粒,转染COS-7细胞。采用胞吞摄取和抗体交联实验,发现nephrin的内吞囊泡与GM1神经节苷脂的十价配体CTxB及podocin囊泡共存;特异性抗体交联促进nephrin与脂筏(lipid raft)标记物CTxB共同聚集于脂筏微区;蔗糖密度梯度离心显示无论是表达nephrin的COS-7细胞还是大鼠肾小球细胞中部分nephrin与脂筏标志物小窝蛋白(caveolin)等均存在于去污剂抵抗膜成分中。结果提示nephrin为脂筏相关蛋白,并且特异抗体交联促进nephrin聚集于脂筏微区。To explore the expressing character of nephrin on cell membrane, constructed plasmid vectors expressing nephrin, podocin. After COS-7cells were transfected, internalization assay and antibody induced crosslinking assay were performed. A colocalization of nephrin and CTxB or podocin were found in COS-7 cells. Antibody induced cross-linking assay showed nephrin colocalized with raft markers. After sucrose floatation gradient centrifugation nephrin floated in detergent resistant membrane fraction together with caveolin and podocin but not transferrin. Results suggest that nephrin is a raft-associated protein and can be recruited by antibody cross-linking.

关 键 词：NEPHRIN 脂筏 抗体交联 

分 类 号：Q25[生物学—细胞生物学]