title: Lipid association, but not the transmembrane domain, is required for tissue factor activity. Substitution of the transmembrane domain with a phosphatidylinositol anchor.
abstract: Full-length tissue factor (263 rTF) and three truncated forms have been expressed in human kidney 293 cells; 1) 243 rTF, which lacks the cytoplasmic tail, is fully functional in the chromogenic assay and has a specific activity comparable with that of the full-length molecule, 263 rTF; 2) 219 rTF, which lacks both the transmembrane and cytoplasmic domains, is not functional; 3) the third variant, referred to as TF-PI, is a fusion protein containing the extracellular domain of TF (amino acids 1-219) fused to the last 37 amino acids of decay-accelerating factor which contain a signal for attachment of a phosphatidylinositol membrane anchor (PI). TF-PI is a membrane-bound protein expressed on the cell surface. The PI anchor restores TF activity lost when the transmembrane domain is deleted from the 219 rTF variant. The ability of the PI anchor to restore activity to 219 rTF clearly demonstrates that while the transmembrane domain is not required for TF activity, lipid association is required.
authors.affiliation: Department of Cardiovascular Research, Genentech, Inc., South San Francisco, California 94080.
authors.forename: L R
authors.initials: LR
authors.lastname: Paborsky
authors.forename: I W
authors.initials: IW
authors.lastname: Caras
authors.forename: K L
authors.initials: KL
authors.lastname: Fisher
authors.forename: C M
authors.initials: CM
authors.lastname: Gorman
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002872:Chromosome Deletion; D005167:Factor VII / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006017:Glycolipids / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols; D006801:Humans; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism*; D010957:Plasmids; D015183:Restriction Mapping; D013925:Thromboplastin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D010716:Phosphatidylinositols; D005167:Factor VII; D013925:Thromboplastin