Energetic Manipulation of Chloroplast Protein Import and the Use of Chemical Cross-Linkers to Map ProteinProtein Interactions
Most chloroplast proteins are synthesized in the cytosol as preproteins with N-terminal cleavable transit peptides and are imported into the organelle through the TOC–TIC translocon system. Import involves a complex set of recognition and membrane translocation steps that ensure the fidelity and unidirectional transport of the polypeptide across the double-membrane chloroplast envelope. To understand the mechanism of import, the molecular interactions and energetics of each step must be defined. Here, we describe the methods for capturing intermediates in the import process through the manipulation of the energy state of chloroplasts, and the use of two different chemical cross-linking approaches to examine the molecular interactions that mediate the import process and to assess the assembly state of the translocons. These approaches can be employed to identify sequential protein–protein interactions, and thereby dissect the pathway and roles of import components during protein import into chloroplasts.
- 植物細(xì)胞滲透勢(shì)的測(cè)定(質(zhì)壁分離法)
- 木栓層
- 鱗片葉
- On-Chip Detection of Low-Molecular-Weight Recombinant Proteins in Plant Crude Extracts by SELDI-TOF MS
- Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic Analysis
- 巢蕨(Neottopteris nidus)
- 球莖
- Arabidopsis Embryogenesis
- Pull-Down Analysis of Interactions Among Jasmonic Acid Core Signaling Proteins
- Suppression Subtractive Hybridization as a Tool to Identify Anthocyanin Metabolism-Related Genes in Apple Skin