title: Charged residues are major determinants of the transmembrane orientation of a signal-anchor sequence.
abstract: Uncleaved signal-anchor sequences of membrane proteins inserted into the endoplasmic reticulum initiate the translocation of either the amino-terminal or the carboxyl-terminal polypeptide segment across the bilayer. Which topology is acquired is not determined by the apolar segment of the signal but rather by the hydrophilic sequences flanking it. To study the role of charged residues in determining the membrane topology, the insertion of mutants of the asialoglycoprotein receptor H1, a single-spanning protein with a cytoplasmic amino terminus, was analyzed in transfected COS-7 cells. When the charged amino acids flanking the hydrophobic signal were mutated to residues of opposite charge, half the polypeptides inserted with the inverted orientation. When, in addition, the amino-terminal domain of the mutant protein was truncated, approximately 90% of the polypeptides acquired the inverted topology. The transmembrane orientation appears to be primarily determined by the charges flanking the signal sequence but is modulated by the domains to be translocated.
authors.affiliation: Department of Biochemistry, University of Basel, Switzerland.
authors.forename: J P
authors.initials: JP
authors.lastname: Beltzer
authors.forename: K
authors.initials: K
authors.lastname: Fiedler
authors.forename: C
authors.initials: C
authors.lastname: Fuhrer
authors.forename: I
authors.initials: I
authors.lastname: Geffen
authors.forename: C
authors.initials: C
authors.lastname: Handschin
authors.forename: H P
authors.initials: HP
authors.lastname: Wessels
authors.forename: M
authors.initials: M
authors.lastname: Spiess
mesh_terms: D000595:Amino Acid Sequence; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D015342:DNA Probes; D008565:Membrane Proteins; D011971:Receptors, Immunologic