Physiology and methane productivity of Methanobacterium thermaggregans

Applied Microbiology and Biotechnology - Accumulation of carbon dioxide (CO2), associated with global temperature rise, and drastically decreasing fossil fuels necessitate the development...     

About the morphological relationships of the sarcoplasmic reticulum in the sole plate area of the frog

In the present investigation the sole plate area of motor end plates of the frog is ultrastructurally examined with different postfixation methods. We concentrated in this case on the proof of the smooth and rough sarcoplasmic reticulum of the sole plate. The relations of the smooth and rough sarcoplasmic reticulum to subsynaptic folds and the local T-system and its connections to diads and triads in the sole plate area are represented. The morphological differences between mammal and frog are pointed out. The possible functions of the sarcoplasmic reticulum in the myofibril-free sarcoplasm are discussed.     

Ultrastructure of the dendritic outer segments of sensory cells in poreless (‘no-pore’) sensilla of insects

Summary Poreless sensilla with inflexible sockets in insects presumably house hygro- and thermoreceptors (type-1, type-2 receptors). The dendritic outer segments of these receptor cells were studied mainly in cryofixed antennae of two species of moth (Antheraea pernyi, A. polyphemus) and one beetle (Aleochara curtula). As a rule two type-1 receptor cells are present. Their dendritic outer segments do not branch. They project into the distal cuticular parts of the sensillum and are in close contact with its four-layered wall. The segments differ in shape and microtubule density. As well, in A. curtula, the microtubules are interconnected by electron-dense material for some distance, thus forming a tubular body-like structure of 1.3 m length. The dendritic outer segment of the single type-2 receptor cell is branched and lamellated. Its lamellae are connected by structures similar to septate junctions, which occupy about 70% of the total surface of the lamellated portion of the dendrite. In tangential sections, the septa appear as parallel strands approximately perpendicular to the long axis of the dendritic segment. The structure of type-1 receptors is discussed with regard to the hypothesis for a mechano-electrical transduction. The possible functions of lamellation and junctional connections in type-2 receptors are discussed.Supported by the Deutsche Forschungsgemeinschaft (SFB 4/G1)     

Biliverdin binds covalently to agrobacterium phytochrome Agp1 via its ring A vinyl side chain

The widely distributed phytochrome photoreceptors carry a bilin chromophore, which is covalently attached to the protein during a lyase reaction. In plant phytochromes, the natural chromophore is coupled by a thioether bond between its ring A ethylidene side chain and a conserved cysteine residue within the so-called GAF domain of the protein. Many bacterial phytochromes carry biliverdin as natural chromophore, which is coupled in a different manner to the protein. In phytochrome Agp1 of Agrobacterium tumefaciens, biliverdin is covalently attached to a cysteine residue close to the N terminus (position 20). By testing different natural and synthetic biliverdin derivatives, it was found that the ring A vinyl side chain is used for chromophore attachment. Only those bilins that have ring A vinyl side chain were covalently attached, whereas bilins with an ethylidene or ethyl side chain were bound in a noncovalent manner. Phycocyanobilin, which belongs to the latter group, was however covalently attached to a mutant in which a cysteine was introduced into the GAF domain of Agp1 (position 249). It is proposed that the regions around positions 20 and 249 are in close contact and contribute both to the chromophore pocket. In competition experiments it was found that phycocyanobilin and biliverdin bind with similar strength to the wild type protein. However, in the V249C mutant, phycocyanobilin bound much more strongly than biliverdin. This finding could explain why during phytochrome evolution in cyanobacteria, the chromophore-binding site swapped from the N terminus into the GAF domain.     

Crystal structure of the IIB(Sor) domain of the sorbose permease from Klebsiella pneumoniae solved to 1.75A resolution

The phosphoenolpyruvate transferase system (PTS) is the major pathway by which bacteria import hexose sugars across the plasma membrane. The PTS transfers a phosphoryl group sequentially via several components from the glycolytic intermediate phosphoenolpyruvate (PEP) to the translocated sugar. It is comprised of the two general proteins enzyme I and HPr, and a sugar-specific enzyme II complex. Sugar translocation is through the membrane domain of the enzyme II complex. The enzyme II complex can belong to one of six families based upon sequence similarity, with the sorbose transporter from Klebsiella pneumoniae a member of the mannose family.The structure of the IIB(Sor) domain was solved to 1.75A resolution by molecular replacement. It has a central core of seven parallel beta-strands surrounded by a total of six alpha-helices. Three helices cover the front face, one the back face with the remaining two capping the central beta-sheet at the top and bottom. The catalytic His15 residue is situated on the surface-exposed loop between strand 1 and helix 1. In addition to the features previously observed in the homologous IIB(Lev) domain from Bacillus subtilis we see new features in the IIB(Sor) structure. First, the catalytic His15 side-chain is fixed in a specific conformation by forming a short hydrogen bond with Asp10, which in turn makes a salt-bridge with Arg8. Second, as observed in other phosphoproteins, an arginine residue (Arg12) is well poised to stabilize a phosphoryl group on His15. Third, we see an Asp/His pair reminiscent of that observed in the IIA(Man) domain from Escherichia coli. Finally, docking of IIA(Man) to IIB(Sor) shows that Arg12 in its current conformation is well positioned to assist the subsequent transfer of the phosphoryl group onto the sugar in line with previous mutagenesis studies.     

Cytochrome c dissociation and release from mitochondria by truncated Bid and ceramide

We have examined the effects of truncated Bid (tBid) and ceramide on mitochondrial membrane integrity and cytochrome c release, using mitochondria with intact outer membranes. While tBid permeabilizes the outer membrane and efficiently stimulates cytochrome c release, digitonin is unable to cause cytochrome c release in the absence of salt. Ceramides did not permeabilize the mitochondrial outer membrane, and stimulated cytochrome c release only in the presence of digitonin. Taken together, these observations support a model for cytochrome c release in which the first step is dissociation from the inner membrane followed by transit across the outer membrane.     

Functional characterization of the interaction between human La and hepatitis B virus RNA

The La protein is a multifunctional RNA-binding protein and has also been suggested to be involved in the stabilization of hepatitis B virus (HBV) RNA. Here we demonstrate that antibodies against the human La protein specifically precipitate HBV RNA from HBV ribonucleoprotein-containing mammalian cell extracts, providing evidence for the association between human La and HBV RNA. Moreover, we report that the turnover of HBV RNA depends on structural features and less on the primary sequence of the La-binding site on the viral RNA. In addition we show that the interaction between human La and HBV RNA in vitro is modulated by accessory factor(s) in a phosphorylation-dependent manner. Taken together these data indicate that both structural features, the composition of La/HBV ribonucleoprotein particles as well as interacting cellular factors, are critical determinants in the regulation of the stability of the HBV RNA.