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Title: Identification of Protein Transport Complexes in the Chloroplastic Envelope Membranes via Chemical Cross-Linking

Journal Article · · Journal of Cell Biology
 [1];  [1];  [1]
  1. Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory
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Transport of cytoplasmically synthesized proteins into chloroplasts uses an import machinery present in the envelope membranes. To identify the components of this machinery and to begin to examine how these components interact during transport, chemical cross-linking was performed on intact chloroplasts containing precursor proteins trapped at a particular stage of transport by ATP limitation. In this work, large crosslinked complexes were observed using three different reversible homobifunctional cross-linkers. Three outer envelope membrane proteins (OEP86, OEP75, and OEP34) and one inner envelope membrane protein (IEP110), previously reported to be involved in protein import, were identified as components of these complexes. In addition to these membrane proteins, a stromal member of the hsp100 family, ClpC, was also present in the complexes. We propose that ClpC functions as a molecular chaperone, cooperating with other components to accomplish the transport of precursor proteins into chloroplasts. We also propose that each envelope membrane contains distinct translocation complexes and that a portion of these interact to form contact sites even in the absence of precursor proteins.

Research Organization:
Michigan State Univ., East Lansing, MI (United States)
Sponsoring Organization:
USDOE Office of Science (SC); National Science Foundation (NSF)
OSTI ID:
1625103
Journal Information:
Journal of Cell Biology, Vol. 136, Issue 5; ISSN 0021-9525
Publisher:
Rockefeller University PressCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (25)

Recognition and envelope translocation of chloroplast preproteins journal August 2005
Characterization of the translocon of the outer envelope of chloroplasts journal February 2003
The Hydrophilic Domain of Tic110, an Inner Envelope Membrane Component of the Chloroplastic Protein Translocation Apparatus, Faces the Stromal Compartment journal June 1998
Common ground for protein translocation: access control for mitochondria and chloroplasts journal December 2010
The Function and Diversity of Plastid Protein Import Pathways: A Multilane GTPase Highway into Plastids: Chloroplast Protein Import journal January 2006
BnaC.Tic40, a plastid inner membrane translocon originating from Brassica oleracea, is essential for tapetal function and microspore development in Brassica napus: Bnac.Tic40 affects tapetal development in Brassica napus journal October 2011
The integration of chloroplast protein targeting with plant developmental and stress responses journal December 2017
Protein import into chloroplasts and its regulation by the ubiquitin-proteasome system journal January 2020
Co-suppression of NbClpC1 and NbClpC2, chaperone subunits in the Clp protease complex, accelerates hypersensitive response and increases disease susceptibility in Nicotiana benthamiana journal June 2019
Plant Hsp100/ClpB-like proteins: poorly-analyzed cousins of yeast ClpB machine journal September 2010
Import of Soluble Proteins into Chloroplasts and Potential Regulatory Mechanisms journal February 2017
The Clp protease system; a central component of the chloroplast protease network journal August 2011
The TIC complex uncovered: The alternative view on the molecular mechanism of protein translocation across the inner envelope membrane of chloroplasts journal September 2015
Protein import into chloroplasts: The Tic complex and its regulation journal June 2010
Molecular chaperone involvement in chloroplast protein import journal February 2013
The chloroplast protein import system: From algae to trees journal February 2013
Protein import into chloroplasts—How chaperones feature into the game journal March 2011
Quantitative Analysis of the Chloroplast Molecular Chaperone ClpC/Hsp93 in Arabidopsis Reveals New Insights into Its Localization, Interaction with the Clp Proteolytic Core, and Functional Importance journal April 2014
Positive Charges Determine the Topology and Functionality of the Transmembrane Domain in the Chloroplastic Outer Envelope Protein Toc34 journal May 1998
Tic20 and Tic22 Are New Components of the Protein Import Apparatus at the Chloroplast Inner Envelope Membrane journal November 1998
The targeting of the atToc159 preprotein receptor to the chloroplast outer membrane is mediated by its GTPase domain and is regulated by GTP journal December 2002
Accumulation and processing of a recombinant protein designed as a cleavable fusion to the endogenous Rubisco LSU protein in Chlamydomonas chloroplast journal March 2009
A J-Like Protein Influences Fatty Acid Composition of Chloroplast Lipids in Arabidopsis journal October 2011
A Protochlorophyllide (Pchlide) a Oxygenase for Plant Viability journal May 2019
Co-Suppression of NbClpC1 and NbClpC2, Encoding Clp Protease Chaperons, Elicits Significant Changes in the Metabolic Profile of Nicotiana benthamiana journal February 2020

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