title: Fusion of sequence elements from non-anchored proteins to generate a fully functional signal for glycophosphatidylinositol membrane anchor attachment.
abstract: Glycophosphatidylinositol (GPI) membrane anchor attachment is directed by a cleavable signal at the COOH terminus of the protein. The complete lack of homology among different GPI-anchored proteins suggests that this signal is of a general nature. Previous analysis of the GPI signal of decay accelerating factor (DAF) suggests that the minimal requirements for GPI attachment are (a) a hydrophobic domain and (b) a cleavage/attachment site consisting of a pair of small residues positioned 10-12 residues NH2-terminal to a hydrophobic domain. As an ultimate test of these rules we constructed four synthetic GPI signals, meeting these requirements but assembled entirely from sequence elements not normally involved in GPI attachment. We show that these synthetic signals are able to direct human growth hormone (hGH), a secreted protein, to the plasma membrane via a GPI anchor. Our results indicate that different hydrophobic sequences, derived from either the prolactin or hGH NH2-terminal signal peptide, can be linked to different cleavage sites via different hydrophilic spacers to produce a functional GPI signal. These data confirm that the only requirements for GPI-anchoring are a pair of small residues positioned 10-12 residues NH2 terminal to a hydrophobic domain, no other structural motifs being necessary.
authors.affiliation: Department of Immunobiology, Genentech, Inc., South San Francisco, California 94080.
authors.forename: P
authors.initials: P
authors.lastname: Moran
authors.forename: I W
authors.initials: IW
authors.lastname: Caras
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D004983:Ethanolamines / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006017:Glycolipids / Q000737:chemistry / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D021382:Protein Sorting Signals / Q000138:chemical synthesis / Q000378:metabolism*; D014162:Transfection; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D004983:Ethanolamines; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D010716:Phosphatidylinositols; D021382:Protein Sorting Signals; D010738:Type C Phospholipases