title: The transit sequence of ferredoxin contains different domains for translocation across the outer and inner membrane of the chloroplast envelope.
abstract: Deletion mutants in the transit sequence of preferredoxin were used in label transfer cross-linking assays to map the interactions of the transit sequence with the import machinery. The deletion mutants gave distinct cross-linking patterns to the Toc and Tic components of the import machinery, consistent with the binding and import properties obtained in in vitro import assays. The cross-linking results revealed two separate properties of the transit peptide: first the presentation of specific binding domains for the initial interaction with outer membrane components, and second the presence of different domains for interaction with the outer and inner membrane components of the transport machinery for full envelope translocation. The N-terminal Delta6-14 deletion blocked import of the precursor at the Toc components, whereas the more internal deletion Delta15-25 blocked import at the Tic components. The information for association with the outer and inner membrane components therefore resides in two separate but partly overlapping domains in the first 25 amino acids of the transit sequence.
authors.affiliation: Department of Molecular Cell Biology, Utrecht University, Padualaan 8, 3584 Utrecht, The Netherlands. W.A.Rensink@bio.uu.nl
authors.forename: W A
authors.initials: WA
authors.lastname: Rensink
authors.forename: D J
authors.initials: DJ
authors.lastname: Schnell
authors.forename: P J
authors.initials: PJ
authors.lastname: Weisbeek
mesh_terms: D000595:Amino Acid Sequence; D002734:Chlorophyll / Q000378:metabolism; D002736:Chloroplasts / Q000378:metabolism*; D003432:Cross-Linking Reagents; D005288:Ferredoxins / Q000737:chemistry* / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D003432:Cross-Linking Reagents; D005288:Ferredoxins; D011498:Protein Precursors; D011994:Recombinant Proteins; D002734:Chlorophyll