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Functional characterization of recombinant chloroplast signal recognition particle

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UNSPECIFIED (2001) Functional characterization of recombinant chloroplast signal recognition particle. JOURNAL OF BIOLOGICAL CHEMISTRY, 276 (30). pp. 27778-27786. ISSN 0021-9258

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Abstract

The signal recognition particle (SRP) is a ubiquitous system for the targeting of membrane and secreted proteins. The chloroplast SRP (cpSRP) is unique among SRPs in that it possesses no RNA and is functional in post-translational as well as co-translational targeting. We have expressed and purified the two components of the Arabidopsis thaliana chloroplast signal recognition particle (cpSRP) involved in post-translational transport: cpSRP54 and the chloroplast-specific protein, cpSRP43. Recombinant cpSRP supports the efficient arc vitro insertion of pea preLhcb1 into isolated thylakoid membranes. Recombinant cpSRP is a stable heterodimer with a molecular mass of similar to 100 kDa as determined by analytical ultracentrifugation, gel filtration analysis, and dynamic light scattering. The interactions of the components of the recombinant heterodimer and pea preLhcb1 were probed using an immobilized peptide library (pepscan) approach. These data confirm two previously reported interactions with the L18 region and the third transmembrane helix of Lhcb1 and suggest that the interface of the cpSRP43 and epSRP54 proteins is involved in substrate binding. Additionally, cpSRP components are shown to recognize peptides from the cleavable, N-terminal chloroplast transit peptide of preLhcb1. The interaction of cpSRP43 with epSRP54 was probed in a similar experiment with a peptide library representing cpSPR54. The C terminus of cpSRP54 is essential for the formation of the stable cpSRP complex and cpSPR43 interacts with distinct regions of the M domain of cpSRP54.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Journal or Publication Title: JOURNAL OF BIOLOGICAL CHEMISTRY
Publisher: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
ISSN: 0021-9258
Date: 27 July 2001
Volume: 276
Number: 30
Number of Pages: 9
Page Range: pp. 27778-27786
Publication Status: Published
URI: http://wrap.warwick.ac.uk/id/eprint/11937

Data sourced from Thomson Reuters' Web of Knowledge

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