title: Multiple determinants direct the orientation of signal-anchor proteins: the topogenic role of the hydrophobic signal domain.
abstract: The orientation of signal-anchor proteins in the endoplasmic reticulum membrane is largely determined by the charged residues flanking the apolar, membrane-spanning domain and is influenced by the folding properties of the NH2-terminal sequence. However, these features are not generally sufficient to ensure a unique topology. The topogenic role of the hydrophobic signal domain was studied in vivo by expressing mutants of the asialoglycoprotein receptor subunit H1 in COS-7 cells. By replacing the 19-residue transmembrane segment of wild-type and mutant H1 by stretches of 7-25 leucine residues, we found that the length and hydrophobicity of the apolar sequence significantly affected protein orientation. Translocation of the NH2 terminus was favored by long, hydrophobic sequences and translocation of the COOH terminus by short ones. The topogenic contributions of the transmembrane domain, the flanking charges, and a hydrophilic NH2-terminal portion were additive. In combination these determinants were sufficient to achieve unique membrane insertion in either orientation.
authors.affiliation: Biozentrum, University of Basel, Switzerland.
authors.forename: J M
authors.initials: JM
authors.lastname: Wahlberg
authors.forename: M
authors.initials: M
authors.lastname: Spiess
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D037263:Asialoglycoprotein Receptor; D001692:Biological Transport; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D006596:Hexosaminidases / Q000494:pharmacology; D007930:Leucine / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D021382:Protein Sorting Signals / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012995:Solubility; D013329:Structure-Activity Relationship; D014357:Trypsin
chemical_list: D037263:Asialoglycoprotein Receptor; D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals; D011956:Receptors, Cell Surface; D006596:Hexosaminidases; D014357:Trypsin; D007930:Leucine