FRET reveals the organization of different receptor-Iigand complexes (polymeric IgA-R and Transferrin-R) in endocytic membranes of polarized MDCK cells

机译：FRET揭示极化的MDCK细胞内吞膜中不同受体-配体复合物（聚合IgA-R和Transferrin-R）的组织

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FRET-based assay has been used to determine the organization of transferrin-receptor bound to holo-transferrin in basolateral endocytic membranes and compare it to the previously characterized clustered organization of polymeric IgA-receptor (plgA-R) bound to plgA-R ligand in apical endocytic membranes. In polarized MDCK-PTR cells, we have internalized holo-transferrin from the basolateral plasma membrane - labeled with donor and acceptor fluorophores. Transferrin-receptor-holo-transferrin complexes were imaged in the basolateral endocytic compartment using FRET confocal laser scanning microscopy in fixed and live MDCK polarized cells. A two-parameter FRET assay demonstrates whether complexes are randomly distributed or clustered: Acceptor's positive impact on E% signifies random distribution; E% being independent of acceptor fluorescence levels indicates clusters. A second parameter for clustering is E% being negatively dependent on D:A ratios. Our results indicating a clustered organization of transferrin-receptor-holo transferrin complexes fit the well-known homodimeric structure of transferrin-receptor.

机译：基于FRET的测定已用于确定基底外侧内吞膜中与全运铁蛋白结合的运铁蛋白受体的组织，并将其与先前表征的与PlgA-R配体结合的聚合IgA受体（plgA-R）的簇状组织进行比较顶端内吞膜。在极化的MDCK-PTR细胞中，我们从基底外侧质膜内化了全运铁蛋白-用供体和受体荧光团标记。使用FRET共聚焦激光扫描显微镜在固定和活的MDCK极化细胞中，在基底外侧内吞区室中对运铁蛋白-受体-全运铁蛋白复合物成像。两参数FRET分析表明复合物是随机分布还是聚集：受体对E％的积极影响表示随机分布； E％独立于受体荧光水平表示簇。聚类的第二个参数是E％负相关于D：A比。我们的结果表明，运铁蛋白-受体-全运铁蛋白复合物的簇状组织符合运铁蛋白-受体的同源二聚体结构。

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来源
《Conference on Multiphoton Microscopy in the Biomedical Sciences IV; 20040125-20040127; San Jose,CA; US》|2004年|P.44-51|共8页
会议地点 San Jose CA(US)
作者
Horst Wallrabe; Margarida Barroso;
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作者单位

University of Virginia, Dep. of Biology, W. M. Keck Center for Cellular Imaging, Gilmer Hall, Charlottesville, VA 22904;

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会议组织
原文格式 PDF
正文语种 eng
中图分类光、激光生物医学;
关键词
transferrin-receptor; transferrin; basolateral; endosome; MDCK; energy transfer efficiency; E; FRET; cluster;

机译：转铁蛋白受体;转铁蛋白;基底外侧;内体; MDCK;能量转移效率; E％; FRET;簇;

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专利

1. Surface Charge of Nanoparticles Determines Their Endocytic and Transcytotic Pathway in Polarized MDCK Cells [J] . Oshrat Harush-Frenkel, Eva Rozentur, Simon Benita Biomacromolecules . 2008,第2期

机译：纳米粒子的表面电荷决定了极化MDCK细胞的内吞途径和跨细胞途径
2. Exocyst requirement for endocytic traffic directed toward the apical and basolateral poles of polarized MDCK cells [J] . Oztan A, Silvis M, Weisz OA, Molecular biology of the cell . 2007,第10期

机译：胞外运输对极化MDCK细胞的顶极和基底外侧极的胞囊需求
3. Exocyst Requirement for Endocytic Traffic Directed Toward the Apical and Basolateral Poles of Polarized MDCK Cells [J] . Asli Oztan, Mark Silvis, Ora A. Weisz, Molecular biology of the cell . 2007,第10期

机译：外囊对内向交通的极化MDCK细胞的顶端和基底外侧极点的要求。
4. FRET reveals the organization of different receptor-Iigand complexes (polymeric IgA-R and Transferrin-R) in endocytic membranes of polarized MDCK cells [C] . Horst Wallrabe, Margarida Barroso, SPIE-The International Society for Optical Engineering, Conference on multiphoton microscopy in the biomedical sciences . 2004

机译：FRET在极化MDCK细胞的内吞膜中揭示了不同受体 - IIGAND复合物（聚合物IgA-R和转移rin-R）的组织
5. Arf6 regulates endocytic trafficking in two examples of polarized cells. [D] . Powelka, Aimee Marie. 2003

机译：Arf6在极化细胞的两个例子中调节内吞运输。
6. Exocyst Requirement for Endocytic Traffic Directed Toward the Apical and Basolateral Poles of Polarized MDCK Cells [O] . Asli Oztan, Mark Silvis, Ora A. Weisz, 2007

机译：外囊对内向交通的极化MDCK细胞的顶端和基底外侧极点的要求。
7. Exocyst Requirement for Endocytic Traffic Directed Toward the Apical and Basolateral Poles of Polarized MDCK Cells [O] . Oztan, Asli, Silvis, Mark, Weisz, Ora A., 2007

机译：外囊对内向交通的极化MDCK细胞的顶端和基底外侧极点的要求。

FRET reveals the organization of different receptor-Iigand complexes (polymeric IgA-R and Transferrin-R) in endocytic membranes of polarized MDCK cells

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