Differential detergent resistance of the apical and basolateral NPPases: relationship with polarized targeting.
Targeting of glycosylphosphatidylinositol-anchored proteins to the apical surface of epithelial cells involves clustering in Triton X-100-resistant membrane microdomains or rafts. The role of these microdomains in sorting transmembrane proteins is more questionable because, unlike glycosylphosphatidylinositol-anchored proteins, apical transmembrane proteins are rather soluble in Triton X-100. They are, however, resistant to milder detergents such as Lubrol WX or Tween 20. It has been proposed that specific membrane microdomains, defined by resistance to these detergents, would carry transmembrane proteins to the apical surface. We have used MDCK cells stably transfected with the apical and basolateral pyrophosphatases/phosphodiesterases, NPP3 and NPP1, to examine the relationship between detergent resistance and apical targeting. The apically expressed wild-type NPP3 was insoluble in Lubrol WX whereas wild-type NPP1, which is expressed basolaterally, was essentially soluble. By using tail mutants and chimeric constructs that combine the cytoplasmic, transmembrane and extracellular domains of NPP1 and NPP3, we show that there is not a strict correlation between detergent resistance and apical targeting. Lubrol resistance is an intrinsic property of NPP3, which is acquired early during the biosynthetic process irrespective of its final destination, and depends on positively charged residues in its cytoplasmic tail.
Delaunay
Jean-Louis
JL
INSERM UMR S 538, Paris, France.
Breton
Michelyne
M
Goding
James W
JW
Trugnan
Germain
G
Maurice
Michèle
M
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology*; D003902:Detergents / Q000737:chemistry*; D004285:Dogs; D021962:Membrane Microdomains / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D017830:Octoxynol / Q000737:chemistry; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry; D011136:Polysorbates / Q000737:chemistry; D021381:Protein Transport / Q000502:physiology; D011755:Pyrophosphatases / Q000235:genetics / Q000378:metabolism*; D051381:Rats
D003902:Detergents; D011136:Polysorbates; C017474:Lubrol WX; D011092:Polyethylene Glycols; D017830:Octoxynol; D010727:Phosphoric Diester Hydrolases; C524442:Enpp3 protein, rat; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; D011755:Pyrophosphatases