caa a22 4500 47712593X CHVBK 20180405111649.0 cr unu---uuuuu 170330e19970901xx s 000 0 eng 10.1023/A:1012167126364 doi (NATIONALLICENCE)springer-10.1023/A:1012167126364 Kinetic Analysis of Transcytosis of Epidermal Growth Factor in Madin-Darby Canine Kidney Epithelial Cells [Elektronische Daten] [Akiko Kozu, Yukio Kato, Yoshihisa Shitara, Manabu Hanano, Yuichi Sugiyama] Purpose. The aim of this study is to clarify the intracellular fate and a rate limiting step in transcytosis of epidermal growth factor (EGF) in Madin-Darby Canine Kidney (MDCK) epithelial cells. Methods. The kinetics of transcytosis of 125I-EGF was investigated. To examine the fate of EGF molecules bound to its receptor on the cell surface, 125I-EGF was prebound to the basal surface at 4°C, followed by extensive washing and subsequent incubation at 37°C in EGF-free medium. Results. Saturable transport of 125I-EGF through the cell monolayer could only be observed from the basal to apical side. Most (~90%) of the EGF molecules bound to the surface receptor are internalized with a half-life of 1−3 min, followed by intracellular degradation with a half-life of 20−50 min. The exocytosis of internalized EGF into the apical medium is much slower with a half-life of 130−250 min. Even when 125I-EGF was incubated with MDCK cells at 37°C and washed with acid to remove cell-surface 125I-EGF, intact 125I-EGF appeared in the basal medium with a half life of 160−170 min. Conclusions. The exocytosis of internalized EGF into the apical medium is a rate limiting step in EGF transcytosis. At least a small amount of internalized EGF is recycled. Plenum Publishing Corporation, 1997 epidermal growth factor nationallicence receptor-mediated endocytosis nationallicence transcytosis nationallicence Madin-Darby Canine Kidney (MDCK) epithelial cell nationallicence Kozu Akiko Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut Kato Yukio Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut Shitara Yoshihisa Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut Hanano Manabu Faculty of Pharmaceutical Sciences, Nihon University, 274, Narashinodai, Funabashi-shi, Chiba, Japan aut Sugiyama Yuichi Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/9(1997-09-01), 1228-1235 0724-8741 14:9<1228 1997 14 11095 https://doi.org/10.1023/A:1012167126364 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012167126364 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kozu Akiko Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Kato Yukio Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Shitara Yoshihisa Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Hanano Manabu Faculty of Pharmaceutical Sciences, Nihon University, 274, Narashinodai, Funabashi-shi, Chiba, Japan aut NATIONALLICENCE 700 1- Sugiyama Yuichi Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/9(1997-09-01), 1228-1235 0724-8741 14:9<1228 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer