title: A few positively charged residues slow movement of a polypeptide chain across the endoplasmic reticulum membrane.
abstract: Many polypeptide chains are translocated across and integrated into the endoplasmic reticulum membrane through protein-conducting channels. During the process, amino acid sequences of translocating polypeptide chains are scanned by the channels and classified to be retained in the membrane or translocated into the lumen. We established an experimental system with which the kinetic effect of each amino acid residue on the polypeptide chain movement can be analyzed with a time resolution of tens of seconds. Positive charges greatly slow movement; only two lysine residues caused a remarkable slow down, and their effects were additive. The lysine residue was more effective than arginine. In contrast, clusters comprising three residues of each of the other 18 amino acids had little effect on chain movement. We also demonstrated that a four lysine cluster can exert the effect after being fully exposed from the ribosome. We concluded that as few as two to three residues of positively charged amino acids can slow the movement of the nascent polypeptide chain across the endoplasmic reticulum membrane. This effect provides a fundamental basis of the topogenic function of positively charged amino acids.
authors.affiliation: Graduate School of Life Science, University of Hyogo , Kouto Ako-gun, Hyogo 678-1297 Japan.
authors.forename: Marifu
authors.initials: M
authors.lastname: Yamagishi
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Onishi
authors.forename: Shotaro
authors.initials: S
authors.lastname: Yoshimura
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Fujita
authors.forename: Kenta
authors.initials: K
authors.lastname: Imai
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Kida
authors.forename: Masao
authors.initials: M
authors.lastname: Sakaguchi
mesh_terms: D000418:Albumins / Q000737:chemistry; D001120:Arginine / Q000737:chemistry; D002474:Cell-Free System; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D012270:Ribosomes / Q000378:metabolism
chemical_list: D000418:Albumins; D010455:Peptides; D001120:Arginine; D008239:Lysine