Expression of active, membrane-bound human placental alkaline phosphatase by transfected simian cells.
Human placental alkaline phosphatase (PALPase) has been transiently expressed in simian (COS) cells by transfection with a eukaryotic expression vector containing the corresponding cDNA. The level of expression of PALPase was high, and it was produced in an enzymatically active form. The bulk of PALPase was associated with the cell membrane as shown by immunocytochemistry and subcellular fractionation studies. The PALPase produced by transfected COS cells, like PALPase in human tissue, was specifically released from the intact cells in a hydrophilic form by phosphatidylinositol-specific phospholipase C and is, therefore, apparently attached to the outer membrane by means of a phosphatidylinositol-glycan. Transfected COS cells appear to be an excellent model for elucidating the mechanism of attachment of this phosphatidylinositol-glycan to a protein moiety.
D000469:Alkaline Phosphatase; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D005347:Fibroblasts / Q000378:metabolism; D058851:GPI-Linked Proteins; D005822:Genetic Vectors; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D004274:DNA, Recombinant; D058851:GPI-Linked Proteins; D007527:Isoenzymes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental