title: An amino-terminal domain containing hydrophobic and hydrophilic sequences binds the signal recognition particle receptor alpha subunit to the beta subunit on the endoplasmic reticulum membrane.
abstract: The signal recognition particle receptor consists of two subunits of 72 kDa (SR alpha) and 30 kDa (SR beta). Assembly of SR alpha on the endoplasmic reticulum membrane can occur independent of the signal recognition particle-mediated translocation pathway. To identify the sequences within SR alpha necessary for membrane binding, a series of amino-terminal and internal deletion mutants was constructed and translated in a cell-free system. In addition, nascent SR alpha polypeptides of varying lengths were generated by cycloheximide treatment of translation reactions. Microsome binding assays performed on these polypeptides revealed a membrane binding domain consisting of the amino-terminal 140 residues of SR alpha. This domain includes the two hydrophobic sequences originally proposed to bind to membranes and a highly charged region not previously implicated in membrane assembly. Furthermore, the domain forms a protease-resistant folding unit that after proteolysis can target and anchor onto microsomes. Extraction of microsomal SR alpha at high pH supplemented with 1 M NaSCN suggests that SR alpha and the membrane binding domain are not integrated in the endoplasmic reticulum membrane. The membrane binding domain is also the major site of tight binding with SR beta, suggesting that SR beta plays a role in the membrane assembly of SR alpha.
authors.affiliation: Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
authors.forename: J C
authors.initials: JC
authors.lastname: Young
authors.forename: J
authors.initials: J
authors.lastname: Ursini
authors.forename: K R
authors.initials: KR
authors.lastname: Legate
authors.forename: J D
authors.initials: JD
authors.lastname: Miller
authors.forename: P
authors.initials: P
authors.lastname: Walter
authors.forename: D W
authors.initials: DW
authors.lastname: Andrews
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D019286:Endopeptidase K; D004721:Endoplasmic Reticulum / Q000737:chemistry* / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D011817:Rabbits; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000378:metabolism; D018000:Receptors, Peptide / Q000737:chemistry* / Q000378:metabolism; D012697:Serine Endopeptidases / Q000494:pharmacology; D018271:Signal Recognition Particle
chemical_list: D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide; D018271:Signal Recognition Particle; C036318:signal peptide receptor; D012697:Serine Endopeptidases; D019286:Endopeptidase K