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Nephrin Clustered in Lipid Raft-associated Microdomain on the Cell Membrane by Antibody Induced Cross-linking
Xiao-Song Qin*, Yong Liu, Wei-Wei Tong, Dan Yue, Jian-Hua Liu, Yan Liu
Department of Laboratory Medicine, Shengjing Hospital of China Medical University, Shenyang 110004, China
Abstract: 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 cross-linking 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.
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