Tracking the transport of E-cadherin to and from the plasma membrane.
The epithelial to mesenchymal transition (EMT) is the breakdown of epithelial cell morphology that gives way to a more mobile, mesenchymal phenotype. Although this process is fundamental to the development of multicellular organisms, it is also a key occurrence in many diseases, including cancers of epithelial origin E-cadherin is a central component of adherens junctions (AJs), which act as structural and signaling hubs in epithelial cells that oppose EMT. The loss of E-cadherin from the plasma membrane is an early indication of EMT and a marker of poor prognosis in many cancers making the trafficking of E-cadherin an area of great interest. Recent work from the authors' laboratory has established the role of type I gamma phosphatidylinositol 4-phosphate 5-kinase (PIPKI gamma) in the trafficking of E-cadherin by studying the surface accessibility of E-cadherin in endocytosis and recycling assays. Additionally, immunofluorescence data demonstrated that cells lacking PIPKI gamma lost E-cadherin at the plasma membrane. The biochemical and microscopic techniques used to investigate the trafficking of E-cadherin are presented herein.
Wagoner
Matthew P
MP
Department of Pharmacology, University of Wisconsin Medical School, Madison, WI, USA.
Ling
Kun
K
Anderson
Richard A
RA
D000818:Animals; D001681:Biological Assay; D001710:Biotin / Q000378:metabolism; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D004285:Dogs; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D008967:Molecular Biology / Q000379:methods*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000172:deficiency / Q000378:metabolism; D021381:Protein Transport; D013194:Staining and Labeling
D015820:Cadherins; D001710:Biotin; D017853:Phosphotransferases (Alcohol Group Acceptor); C020077:1-phosphatidylinositol-4-phosphate 5-kinase