The Transit Sequence of Ferredoxin Contains Different Domains for Translocation across the Outer and Inner Membrane of the Chloroplast Envelope

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, consi...

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Bibliographic Details
Published in:The Journal of biological chemistry 2000-04, Vol.275 (14), p.10265-10271
Main Authors: Rensink, Willem Albert, Schnell, Danny J., Weisbeek, Peter J.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
binding
binding proteins
cell membranes
Chlorophyll - metabolism
chloroplasts
Chloroplasts - metabolism
Cross-Linking Reagents
crosslinking
deletions
Ferredoxin
Ferredoxins - chemistry
Ferredoxins - metabolism
Intracellular Membranes - metabolism
Kinetics
Molecular Sequence Data
Mutagenesis
mutants
Protein Precursors - chemistry
Protein Precursors - metabolism
protein transport
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Sequence Deletion
signal peptide
Silene latifolia subsp. alba
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Summary: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 Δ6–14 deletion blocked import of the precursor at the Toc components, whereas the more internal deletion Δ15–25 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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.275.14.10265