Proteomic Analysis of the Proplastid Envelope Membrane Provides Novel Insights into Small Molecule and Protein Transport across Proplastid Membranes

Bräutigam A, Weber APM. Proteomic Analysis of the Proplastid Envelope Membrane Provides Novel Insights into Small Molecule and Protein Transport across Proplastid Membranes. Molecular Plant. 2009;2(6):1247-1261.Proplastids are undifferentiated plastids of meristematic tissues that synthesize amino acids for protein synthesis, fatty acids for membrane lipid production, and purines and pyrimidines for DNA and RNA synthesis. Unlike chloroplasts, proplastids depend on supply, with reducing power, energy, and precursor metabolites from the remainder of the cell. Comparing proplastid and chloroplast envelope proteomes and the corresponding transcriptomes of leaves and shoot apex revealed a clearly distinct composition of the proplastid envelope. It is geared towards import of metabolic precursors and export of product metabolites for the rapidly dividing cell. The analysis also suggested a new role for the triosephosphate translocator in meristematic tissues, identified the route of organic nitrogen import into proplastids, and detected an adenine nucleotide exporter. The protein import complex contains the import receptors Toc120 and Toc132 and lacks the redox sensing complex subunits of Tic32, Tic55, and Tic62, which mirrors the expression patterns of the corresponding genes in leaves and the shoot apex. We further show that the protein composition of the internal membrane system is similar to etioplasts, as it is dominated by the ATP synthase complex and thus remarkably differs from that of chloroplast thylakoids

The role of membrane transport in metabolic engineering of plant primary metabolism

Weber APM, Bräutigam A. The role of membrane transport in metabolic engineering of plant primary metabolism. Current Opinion in Biotechnology. 2013;24(2):256-262.Plant cells are highly compartmentalized and so is their metabolism. Most metabolic pathways are distributed across several cellular compartments, which requires the activities of membrane transporters to catalyze the flux of precursors, intermediates, and end products between compartments. Metabolites such as sucrose and amino acids have to be transported between cells and tissues to supply, for example, metabolism in developing seeds or fruits with precursors and energy. Thus, rational engineering of plant primary metabolism requires a detailed and molecular understanding of the membrane transporters. This knowledge however still lags behind that of soluble enzymes. Recent advances include the molecular identification of pyruvate transporters at the chloroplast and mitochondrial membranes and of a new class of transporters called SWEET that are involved in the release of sugars to the apoplast

The Metabolite Transporters of the Plastid Envelope: An Update

The engulfment of a photoautotrophic cyanobacterium by a primitive mitochondria-bearing eukaryote traces back to more than 1.2 billion years ago. This single endosymbiotic event not only provided the early petroalgae with the metabolic capacity to perform oxygenic photosynthesis, but also introduced a plethora of other metabolic routes ranging from fatty acids and amino acids biosynthesis, nitrogen and sulfur assimilation to secondary compounds synthesis. This implicated the integration and coordination of the newly acquired metabolic entity with the host metabolism. The interface between the host cytosol and the plastidic stroma became of crucial importance in sorting precursors and products between the plastid and other cellular compartments. The plastid envelope membranes fulfill different tasks: they perform important metabolic functions, as they are involved in the synthesis of carotenoids, chlorophylls, and galactolipids. In addition, since most genes of cyanobacterial origin have been transferred to the nucleus, plastidial proteins encoded by nuclear genes are post-translationally transported across the envelopes through the TIC–TOC import machinery. Most importantly, chloroplasts supply the photoautotrophic cell with photosynthates in form of reduced carbon. The innermost bilayer of the plastidic envelope represents the permeability barrier for the metabolites involved in the carbon cycle and is literally stuffed with transporter proteins facilitating their transfer. The intracellular metabolite transporters consist of polytopic proteins containing membrane spans usually in the number of four or more α-helices. Phylogenetic analyses revealed that connecting the plastid with the host metabolism was mainly a process driven by the host cell. In Arabidopsis, 58% of the metabolite transporters are of host origin, whereas only 12% are attributable to the cyanobacterial endosymbiont. This review focuses on the metabolite transporters of the inner envelope membrane of plastids, in particular the electrochemical potential-driven class of transporters. Recent advances in elucidating the plastidial complement of metabolite transporters are provided, with an update on phylogenetic relationship of selected proteins

Ultrafast Dynamics of Carrier Multiplication in Quantum Dots

A quantum-kinetic approach to the ultrafast dynamics of carrier multiplication in semiconductor quantum dots is presented. We investigate the underlying dynamics in the electronic subband occupations and the time-resolved optical emission spectrum, focusing on the interplay between the light-matter and the Coulomb interaction. We find a transition between qualitatively differing behaviors of carrier multiplication, which is controlled by the ratio of the interaction induced time scale and the pulse duration of the exciting light pulse. On short time scales, i.e., before intra-band relaxation, this opens the possibility of detecting carrier multiplication without refering to measurements of (multi-)exciton lifetimes.Comment: 12 pages, 7 figures, submitte

Lie Symmetry Analysis for Cosserat Rods

We consider a subsystem of the Special Cosserat Theory of Rods and construct an explicit form of its solution that depends on three arbitrary functions in (s,t) and three arbitrary functions in t. Assuming analyticity of the arbitrary functions in a domain under consideration, we prove that the obtained solution is analytic and general. The Special Cosserat Theory of Rods describes the dynamic equilibrium of 1-dimensional continua, i.e. slender structures like fibers, by means of a system of partial differential equations.Comment: 12 Pages, 1 Figur

Transcriptional profiling of Arabidopsis heat shock proteins and transcription factors reveals extensive overlap between heat and non-heat stress response pathways

<p>Abstract</p> <p>Background</p> <p>The heat shock response of <it>Arabidopsis thaliana </it>is dependent upon a complex regulatory network involving twenty-one known transcription factors and four heat shock protein families. It is known that heat shock proteins (Hsps) and transcription factors (Hsfs) are involved in cellular response to various forms of stress besides heat. However, the role of Hsps and Hsfs under cold and non-thermal stress conditions is not well understood, and it is unclear which types of stress interact least and most strongly with Hsp and Hsf response pathways. To address this issue, we have analyzed transcriptional response profiles of <it>Arabidopsis </it>Hsfs and Hsps to a range of abiotic and biotic stress treatments (heat, cold, osmotic stress, salt, drought, genotoxic stress, ultraviolet light, oxidative stress, wounding, and pathogen infection) in both above and below-ground plant tissues.</p> <p>Results</p> <p>All stress treatments interact with Hsf and Hsp response pathways to varying extents, suggesting considerable cross-talk between heat and non-heat stress regulatory networks. In general, Hsf and Hsp expression was strongly induced by heat, cold, salt, and osmotic stress, while other types of stress exhibited family or tissue-specific response patterns. With respect to the Hsp20 protein family, for instance, large expression responses occurred under all types of stress, with striking similarity among expression response profiles. Several genes belonging to the Hsp20, Hsp70 and Hsp100 families were specifically upregulated twelve hours after wounding in root tissue, and exhibited a parallel expression response pattern during recovery from heat stress. Among all Hsf and Hsp families, large expression responses occurred under ultraviolet-B light stress in aerial tissue (shoots) but not subterranean tissue (roots).</p> <p>Conclusion</p> <p>Our findings show that Hsf and Hsp family member genes represent an interaction point between multiple stress response pathways, and therefore warrant functional analysis under conditions apart from heat shock treatment. In addition, our analysis revealed several family and tissue-specific heat shock gene expression patterns that have not been previously described. These results have implications regarding the molecular basis of cross-tolerance in plant species, and raise new questions to be pursued in future experimental studies of the <it>Arabidopsis </it>heat shock response network.</p

High-throughput colorimetric method for the parallel assay of glyoxylic acid and ammonium in a single extract

Bräutigam A, Gagneul D, Weber APM. High-throughput colorimetric method for the parallel assay of glyoxylic acid and ammonium in a single extract. Analytical Biochemistry. 2007;362(1):151-153

Comparative Proteomics of Chloroplast Envelopes from C-3 and C-4 Plants Reveals Specific Adaptations of the Plastid Envelope to C-4 Photosynthesis and Candidate Proteins Required for Maintaining C-4 Metabolite Fluxes (vol 148, pg 568, 2008)

Bräutigam A, Hoffmann-Benning S, Weber APM. Comparative Proteomics of Chloroplast Envelopes from C-3 and C-4 Plants Reveals Specific Adaptations of the Plastid Envelope to C-4 Photosynthesis and Candidate Proteins Required for Maintaining C-4 Metabolite Fluxes (vol 148, pg 568, 2008). Plant Physiology. 2008;148(3):1734

The Plastid Outer Envelope – A Highly Dynamic Interface between Plastid and Cytoplasm

Plastids are the defining organelles of all photosynthetic eukaryotes. They are the site of photosynthesis and of a large number of other essential metabolic pathways, such as fatty acid and amino acid biosyntheses, sulfur and nitrogen assimilation, and aromatic and terpenoid compound production, to mention only a few examples. The metabolism of plastids is heavily intertwined and connected with that of the surrounding cytosol, thus causing massive traffic of metabolic precursors, intermediates, and products. Two layers of biological membranes that are called the inner (IE) and the outer (OE) plastid envelope membranes bound the plastids of Archaeplastida. While the IE is generally accepted as the osmo-regulatory barrier between cytosol and stroma, the OE was considered to represent an unspecific molecular sieve, permeable for molecules of up to 10 kDa. However, after the discovery of small substrate specific pores in the OE, this view has come under scrutiny. In addition to controlling metabolic fluxes between plastid and cytosol, the OE is also crucial for protein import into the chloroplast. It contains the receptors and translocation channel of the TOC complex that is required for the canonical post-translational import of nuclear-encoded, plastid-targeted proteins. Further, the OE is a metabolically active compartment of the chloroplast, being involved in, e.g., fatty acid metabolism and membrane lipid production. Also, recent findings hint on the OE as a defense platform against several biotic and abiotic stress conditions, such as cold acclimation, freezing tolerance, and phosphate deprivation. Moreover, dynamic non-covalent interactions between the OE and the endomembrane system are thought to play important roles in lipid and non-canonical protein trafficking between plastid and endoplasmic reticulum. While proteomics and bioinformatics has provided us with comprehensive but still incomplete information on proteins localized in the plastid IE, the stroma, and the thylakoids, our knowledge of the protein composition of the plastid OE is far from complete. In this article, we report on the recent progress in discovering novel OE proteins to draw a conclusive picture of the OE. A “parts list” of the plastid OE will be presented, using data generated by proteomics of plastids isolated from various plant sources