Fabrication of a dual substrate display to test roles of cell adhesion proteins in vesicle targeting to plasma membrane domains.
While much is known of the molecular machinery involved in protein sorting during exocytosis, less is known about the spatial regulation of exocytosis at the plasma membrane (PM). This study outlines a novel method, dual substrate display, used to formally test the hypothesis that E-cadherin-mediated adhesion directs basolateral vesicle exocytosis to specific sites at the PM. We show that vesicles containing the basolateral marker protein VSV-G preferentially target to sites of adhesion to E-cadherin rather than collagen VI or a control peptide. These results support the hypothesis that E-cadherin adhesion initiates signaling at the PM resulting in targeted sites for exocytosis.
Hunt
Stephen J
SJ
Department of Biological Sciences, Stanford University, James H. Clark Center, 318 Campus Drive, Room E200, Stanford, CA 94305-5430, United States.
Nelson
W James
WJ
D000818:Animals; D001692:Biological Transport; D015820:Cadherins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D002232:Carbocyanines / Q000737:chemistry; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D022162:Cytoplasmic Vesicles / Q000378:metabolism*; D005089:Exocytosis; D005456:Fluorescent Dyes / Q000737:chemistry; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism