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Binding of chloroplast signal recognition particle to a thylakoid membrane protein substrate in aqueous solution and delineation of the cpSRP43–substrate interaction domain

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Cain, Peter, Holdermann, Iris, Sinning, Irmgard, Johnson, Arthur E. and Robinson, Colin. (2011) Binding of chloroplast signal recognition particle to a thylakoid membrane protein substrate in aqueous solution and delineation of the cpSRP43–substrate interaction domain. Biochemical Journal, Vol.437 (No.1). pp. 149-155. ISSN 0264-6021

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Official URL: http://dx.doi.org/10.1042/BJ20110270

Abstract

A cpSRP [chloroplast SRP (signal recognition particle)] comprising cpSRP54 and cpSRP43 subunits mediates the insertion of light-harvesting proteins into the thylakoid membrane. We dissected its interaction with a full-length membrane protein substrate in aqueous solution by insertion of site-specific photoactivatable cross-linkers into in vitro-synthesized Lhcb1 (major light-harvesting chlorophyll-binding protein of photosystem II). We show that Lhcb1 residues 166-176 cross-link specifically to the cpSRP43 subunit. Some cross-link positions within Lhcb1 are in the 'L18' peptide required for targeting of cpSRP substrates, whereas other cross-linking positions define a new targeting signal in the third transmembrane span. Lhcb1 was not found to cross-link to cpSRP54 at any position, and cross-linking to cpSRP43 is unaffected by the absence of cpSRP54. cpSRP43 thus effectively binds substrates autonomously, and its ability to independently bind an extended 20+-residue substrate region highlights a major difference with other SRP types where the SRP54 subunit binds to hydrophobic target sequences. The results also show that cpSRP43 can bind to a hydrophobic, three-membrane span, substrate in aqueous solution, presumably reflecting a role for cpSRP in the chloroplast stroma. This mode of action, and the specificity of the cpSRP43 substrate interaction, may be associated with cpSRP's unique post-translational mode of action.

Item Type:

Journal Article

Subjects:

Q Science > QK Botany

Divisions:

Faculty of Science > Life Sciences (2010- )

Library of Congress Subject Headings (LCSH):

Chloroplasts -- Physiology, Plant proteins

Journal or Publication Title:

Biochemical Journal

Publisher:

Portland Press

ISSN:

0264-6021

Official Date:

1 July 2011

Dates:

Date	Event
1 July 2011	Published

Volume:

Vol.437

Number:

No.1

Page Range:

pp. 149-155

Identifier:

10.1042/BJ20110270

Status:

Peer Reviewed

Publication Status:

Published

Access rights to Published version:

Restricted or Subscription Access

Funder:

Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), National Institutes of Health (U.S.) (NIH), Robert A. Welch Foundation

Grant number:

GM26494 (NIH), BE-0017 (Robert A. Welch Foundation)

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