Absence of direct delivery for single transmembrane apical proteins or their "Secretory" forms in polarized hepatic cells.
The absence of a direct route to the apical plasma membrane (PM) for single transmembrane domain (TMD) proteins in polarized hepatic cells has been inferred but never directly demonstrated. The genes encoding three pairs of apical PM proteins, whose extracellular domains are targeted exclusively to the apical milieu in Madin-Darby canine kidney cells, were packaged into recombinant adenovirus and delivered to WIF-B cells in vitro and liver hepatocytes in vivo. By immunofluorescence and pulse-chase metabolic labeling, we found that the soluble constructs were overwhelmingly secreted into the basolateral milieu, which in vivo is the blood and in vitro is the culture medium. The full-length proteins were first delivered to the basolateral surface but then concentrated in the apical PM. Our results imply that hepatic cells lack trans-Golgi network (TGN)-based machinery for directly sorting single transmembrane domain apical proteins and raise interesting questions about current models of PM protein sorting in polarized and nonpolarized cells.
Bastaki
M
M
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Braiterman
L T
LT
Johns
D C
DC
Chen
Y-H
YH
Hubbard
A L
AL
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001646:Bile / Q000737:chemistry; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D016764:Cell Polarity / Q000502:physiology*; D003470:Culture Media / Q000032:analysis; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism*; D007700:Kinetics; D008163:Luminescent Measurements; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D011994:Recombinant Proteins / Q000097:blood / Q000378:metabolism; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D003470:Culture Media; D008565:Membrane Proteins; D011994:Recombinant Proteins