Surfactant proteins are well known from the human lung where they are responsible for the stability and flexibility of the pulmonary surfactant system. They are able to influence the surface tension of the gas–liquid interface specifically by directly interacting with single lipids. This work describes the generation of reliable protein structure models to support the experimental characterization of two novel putative surfactant proteins called SP-G and SP-H. The obtained protein models were complemented by predicted posttranslational modifications and placed in a lipid model system mimicking the pulmonary surface. Molecular dynamics simulations of these protein-lipid systems showed the stability of the protein models and the formation of interactions between protein surface and lipid head groups on an atomic scale. Thereby, interaction interface and strength seem to be dependent on orientation and posttranslational modification of the protein. The here presented modeling was fundamental for experimental localization studies and the simulations showed that SP-G and SP-H are theoretically able to interact with lipid systems and thus are members of the surfactant protein family.

Leaf senescence is the final developmental stage of a leaf. The progression of barley primary leaf senescence was followed by measuring the senescence‐specific decrease in chlorophyll content and photosystem II efficiency. In order to isolate novel factors involved in leaf senescence, a differential display approach with mRNA populations from young and senescing primary barley leaves was applied. In this approach, 90 senescence up‐regulated cDNAs were identified. Nine of these clones were, after sequence analyses, further characterized. The senescence‐associated expression was confirmed by Northern analyses or quantitative RealTime‐PCR. In addition, involvement of the phytohormones ethylene and abscisic acid in regulation of these nine novel senescence‐induced cDNA fragments was investigated. Two cDNA clones showed homologies to genes with a putative regulatory function. Two clones possessed high homologies to barley retroelements, and five clones may be involved in degradation or transport processes. One of these genes was further analysed. It encodes an ADP ribosylation factor 1‐like protein (HvARF1) and includes sequence motifs representing a myristoylation site and four typical and well conserved ARF‐like protein domains. The localization of the protein was investigated by confocal laser scanning microscopy of onion epidermal cells after particle bombardment with chimeric HvARF1‐GFP constructs. Possible physiological roles of these nine novel SAGs during barley leaf senescence are discussed.

Phytohormones are not only instrumental in regulating developmental processes in plants but also play important roles for the plant's responses to biotic and abiotic stresses. In particular, abscisic acid, ethylene, jasmonic acid, and salicylic acid have been shown to possess crucial functions in mediating or orchestrating stress responses in plants. Here, we review the role of salicylic acid and jasmonic acid in pathogen defence responses with special emphasis on their function in the solanaceous plant potato.

Among the plant hormones jasmonic acid and related derivatives are known to mediate stress responses and several developmental processes. Biosynthesis, regulation, and metabolism of jasmonic acid in Arabidopsis thaliana are reviewed, including properties of mutants of jasmonate biosynthesis. The individual signalling properties of several jasmonates are described.

4-Hydroxybenzoate oligoprenyltransferase of E. coli, encoded in the gene ubiA, is an important key enzyme in the biosynthetic pathway to ubiquinone. It catalyzes the prenylation of 4-hydroxybenzoic acid in position 3 using an oligoprenyl diphosphate as a second substrate. Up to now, no X-ray structure of this oligoprenyltransferase or any structurally related enzyme is known. Knowledge of the tertiary structure and possible active sites is, however, essential for understanding the catalysis mechanism and the substrate specificity.With homology modeling techniques, secondary structure prediction tools, molecular dynamics simulations, and energy optimizations, a model with two putative active sites could be created and refined. One active site selected to be the most likely one for the docking of oligoprenyl diphosphate and 4-hydroxybenzoic acid is located near the N-terminus of the enzyme. It is widely accepted that residues forming an active site are usually evolutionary conserved within a family of enzymes. Multiple alignments of a multitude of related proteins clearly showed 100% conservation of the amino acid residues that form the first putative active site and therefore strongly support this hypothesis. However, an additional highly conserved region in the amino acid sequence of the ubiA enzyme could be detected, which also can be considered a putative (or rudimentary) active site. This site is characterized by a high sequence similarity to the aforementioned site and may give some hints regarding the evolutionary origin of the ubiA enzyme.Semiempirical quantum mechanical PM3 calculations have been performed to investigate the thermodynamics and kinetics of the catalysis mechanism. These results suggest a near SN1 mechanism for the cleavage of the diphosphate ion from the isoprenyl unit. The 4-hydroxybenzoic acid interestingly appears not to be activated as benzoate anion but rather as phenolate anion to allow attack of the isoprenyl cation to the phenolate, which appeared to be the rate limiting step of the whole process according to our quantum chemical calculations. Our models are a basis for developing inhibitors of this enzyme, which is crucial for bacterial aerobic metabolism.

Transport processes between plant and fungal cells are key elements in arbuscular mycorrhiza (AM), where H+‐ATPases are considered to be involved in active uptake of nutrients from the symbiotic interface. Genes encoding H+‐ATPases were identified in the genome of Medicago truncatula and three cDNA fragments of the H+‐ATPase gene family (Mtha 1 ‐ 3) were obtained by RT‐PCR using RNA from M. truncatula mycorrhizal roots as template. While Mtha 2 and Mtha 3 appeared to be constitutively expressed in roots and unaffected by AM development, transcripts of Mtha 1 could only be detected in AM tissues and not in controls. Further analyses by RT‐PCR revealed that Mtha 1 transcripts are not detectable in shoots and phosphate availability did not affect RNA accumulation of the gene. Localization of transcripts by in situ hybridization on AM tissues showed that Mtha 1 RNA accumulates only in cells containing fungal arbuscules. This is the first report of arbuscule‐specific induced expression of a plant H+‐ATPase gene in mycorrhizal tissues.

Brassinosteroids are a class of steroidal phytohormones with high growth-promoting properties. The preferred side-chain conformations of 10 brassinosteroids were determined by means of detailed NMR investigations and molecular-modeling studies. Vacuum conformations obtained by simulated annealing calculations and Boltzmann statistical analysis were compared with solution conformations derived from NOE experiments and molecular dynamic simulations, and with X-ray structures. In general, results from simulated annealing calculations and NMR-supported molecular dynamics simulations are in good agreement. For some of the compounds investigated the conformation was less well-defined at the end of the side-chain. It could be shown that the energetically most favorable and most probable conformations also include the conformations obtained by NMR supported molecular-dynamics calculations and by X-ray analysis. For the most bioactive compound brassinolide (1) the majority of conformations show a side-chain bent towards the β-face of the steroid skeleton, whereas for the less bioactive brassinosteroids, conformations with straight side-chains or side-chains bent towards the α-face are more frequent.

Conventional and analytical electron microscopy (EDX, ESI, EELS) were used to investigate the silicon accumulation, the chemical nature of the Si deposits and their formation in three species of monocotyledons. In Deschampsia , in particular parts of the outer epidermal cell wall silicon is accumulated as silicic acid. Electron dense, needle‐shaped crystals in the vacuoles of epidermal cells and in the intercellular spaces were also identified as silicic acid. In xylem parenchyma cells, silicon is accumulated as SiO2, which is formed from Sn silicate. In Festuca , crystal‐like deposits of SiO2 occur on the epidermal surface, in the epidermal and parenchyma cell walls, and in vacuoles of bundle sheath cells. Often the deposits disturb the cell walls and penetrate the envelope of plastids and mitochondria. The crystal‐like SiO2 deposits originate from Sn silicate. In the pericarp of ripe nuts of Schoenus , no stainable cell wall components are detected. The inner part of the pericarp consists of silicic acid, while in the outer regions small clusters of silicic acid are embedded in a matrix of SiO2. The silicic acid deposits show an unusual, layered structure, typical for lepidoic silicic acids, which consist of two‐dimensional crystals lying one above the other.

Treatment of barley leaf segments with jasmonic acid methyl ester (JM) leads to the accumulation of a set of newly formed abundant proteins. Among them, the most abun dant protein exhibits a molecular mass of 23 kDa (JIP‐23). Here, data are presented on the occurrence and expression of the lIP‐23 genes in different cultivars of Hordeum vulgare . Southern blot analysis of 80 cultivars revealed the occurrence of 2 to 4 genes coding for JIP‐23 in all cultivars. By means of Northern blot and immunoblot analysis it is shown that some cultivars lack the ex pression of jip‐23 upon treatment of primary leaves with JM as well as upon stress performed by incubation with 1 M sorbitol solution. During germination, however, all tested cultivars ex hibited developmental expression of jip‐23 . The results are dis cussed in terms of possible functions of JIP‐23 in barley.

Cathepsin H is involved in intracellular protein degradation and is implicated in a variety of physiological processes such as proenzyme activation, enzyme inactivation, hormone maturation, tissue remodeling, and bone matrix resorption. A model of the tertiary structure of the human lysosomal cysteine protease cathepsin H was constructed. The protein structure was built from its amino acid sequence and its homology to papain, actinidin, and cathepsin L for which crystallographic co-ordinates are available. The model was generated using the COMPOSER module of SYBYL.The position and interaction behavior of the so called mini-chain, the octapeptide EPQNCSAT, to the active-site cleft of cathepsin H could be determined by docking studies. Refinement was achieved through interactive visual and algorithmic analysis and minimization with the TRIPOS force field. The model was found to correlate with observed empirical data regarding ligand specificity. The model defines possible steric, hydrophobic, and electrostatic interactions. We anticipate that the model will serve as a tool to understand substrate specificity and may be used for the development of new specific ligands.