title: Positive charges of translocating polypeptide chain retrieve an upstream marginal hydrophobic segment from the endoplasmic reticulum lumen to the translocon.
abstract: Positively charged amino acid residues are well recognized topology determinants of membrane proteins. They contribute to the stop-translocation of a polypeptide translocating through the translocon and to determine the orientation of signal sequences penetrating the membrane. Here we analyzed the function of these positively charged residues during stop-translocation in vitro. Surprisingly, the positive charges facilitated membrane spanning of a marginally hydrophobic segment, even when separated from the hydrophobic segment by 70 residues. In this case, the hydrophobic segment was exposed to the lumen, and then the downstream positive charges triggered the segment to slide back into the membrane. The marginally hydrophobic segment spanned the membrane, but maintained access to the water environment. The positive charges not only fix the hydrophobic segment in the membrane at its flanking position, but also have a much more dynamic action than previously realized.
authors.affiliation: Graduate School of Life Science, University of Hyogo, Kouto Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Fujita
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Kida
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Hagiwara
authors.forename: Fumiko
authors.initials: F
authors.lastname: Morimoto
authors.forename: Masao
authors.initials: M
authors.lastname: Sakaguchi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008954:Models, Biological; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D014867:Water
chemical_list: D000596:Amino Acids; D010455:Peptides; D021382:Protein Sorting Signals; D014867:Water