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Analysis of Protein Import into Chloroplasts Isolated from Stressed Plants
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Analysis of starch metabolism in chloroplasts

Carmen Hostettler1, Katharina Kölling, Diana Santelia

  • 1Department of Biology, ETH Zurich, Zurich, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)
|August 25, 2011
PubMed

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Protein Transport to the Stroma01:24

Protein Transport to the Stroma

Chloroplasts are triple membrane structures with an outer membrane, an inner membrane, and a thylakoid membrane, each containing distinct metabolite transporters, membrane translocons, and enzymes. Appropriate sorting and translocating these proteins to their correct membrane systems is essential for chloroplast function.
Protein complexes called the translocon of the outer chloroplast membrane or TOC complex, and the translocon of the inner chloroplast membrane or TIC complex mediate the...
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The Anatomy of Chloroplasts

Green algae and plants, including green stems and unripe fruit, harbor specialized organelles called chloroplasts to carry out photosynthesis. They coordinate both stages of photosynthesis — the light-dependent reactions and the light-independent reactions. The light-dependent reactions use sunlight to release oxygen and produce chemical energy in the form of ATP and NADPH, and the light-independent reactions capture CO2 and use ATP and NADPH to produce sugar.
Structure of Chloroplasts
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Anatomy of Chloroplasts01:07

Anatomy of Chloroplasts

Green algae and plants, including green stems and unripe fruit, harbor chloroplasts—the vital organelles where photosynthesis takes place. In plants, the highest density of chloroplasts is found in the mesophyll cells of leaves.
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Protein Transport to the Inner Chloroplast Membrane

Proteins targeted to the inner chloroplast membrane, or plastid proteins, are transported by two general pathways: the stop-transfer and the re-insertion or post-import pathways. Most plastid proteins carry N-terminal transit sequences and internal import sequences targeting it to the specific chloroplast subcompartment. Proteins targeted by the stop-transfer pathway have internal hydrophobic sequences that inhibit their translocation into the stroma. As a result, these precursors are arrested...
Protein Transport to the Outer Chloroplast Membrane01:11

Protein Transport to the Outer Chloroplast Membrane

Chloroplast outer membrane proteins encoded by the nucleus are synthesized in the cytosol. Soon after synthesis, they bind cytosolic factors such as 14-3-3 protein and the Hsp70 chaperones that keep these precursors in an unfolded state until their translocation.
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Thylakoids are membrane-bound sac-like structures within the chloroplast that serve as sites for photosynthesis. Thylakoid lumen contains many electron transport proteins and is enclosed by a thylakoid membrane rich in the light-harvesting complex. Proteins targeted to the thylakoids are transported as precursors and are sorted by the general TOC/TIC import pathway. Once the precursor reaches the stroma, stromal processing peptidases remove their transit signal and expose thylakoid signal...