Surface properties of fibrinogen and fibrin

By contact angle measurements on layers of fibrinogen and fibrin, it can be shown that the transformation from fibrinogen to fibrin is accompanied by a change in surface properties from very hydrophilic (fibrinogen) to moderately but definitely hydrophobic (fibrin). It is also shown that, contrary to serum albumin and gamma globulin, fibrinogen does not become more hydrophobic upon drying.     

Soluble and membrane-bound ferrisiderophore reductases of Escherichia coli K-12

After uptake of microbial ferrisiderophores, iron is assumed to be released by reduction. Two ferrisiderophore-reductase activities were identified in Escherichia coli K-12. They differed in cellular location, susceptibility to amytal, and competition between oxygen and ferrichrome-iron(III) reduction. The ferrisiderophore reductase associated with the 40,000×g sediment (membrane-bound enzyme) was inhibited by 10 mM amytal in contrast to the ferrisiderophore reductase present in the 100,000×g supernatant (soluble enzyme). Reduction by the membrane-bound enzyme followed sigmoid kinetics, but was biphasic in the case of the soluble enzyme. The soluble reductase could be assigned to a protein consisting of a single polypeptide of M _r 26000. Reduction of iron(III) by the purified enzyme depended on the addition of NADH or NADPH which were equally active reductants. The cofactor FMN and to a lesser degree FAD stimulated the reaction. Substrate specificity of the soluble reductase was low. In addition to the hydroxamate siderophores arthrobactin, schizokinen, fusigen, aerobactin, ferrichrome, ferrioxamine B, coprogen, and ferrichrome A, the iron(III) complexes of synthetic catecholates, dihydroxy benzoic acid, and dicitrate, as well as carrier-free iron(III) were accepted as substrates. Both ferrisiderophore reductases were not controlled by the fur regulatory system and were not suppressed by anaerobic growth.Abbreviations DHB dihydroxybenzoic acid - MECAM 1,3,5-N,N,N-tris-(2,3-dihydroxybenzoyl)-triamino-methylbenzene - MECAMS 2,3-dihydroxy-5-sulfonyl-derivative of MECAM     

Oxidation of glycerol,lactate, and propionate by Propionibacterium freudenreichii in a poised-potential amperometric culture system

Growth of Propionibacterium freudenreichii was studied with glycerol, lactate, and propionate as energy sources and a three-electrode poised-potential amperometric electrode system with hexacyanoferrate (III) as mediator. In batch culture experiments with glycerol and lactate as substrates, hexacyanoferrate (III) was completely reduced. Growth yields increased and the fermentation patterns were shifted towards higher acetate formation with increasing hexacyanoferrate (III) concentrations (0.25–8.0 mM). In experiments with regulated electrodes, glycerol, lactate, and propionate were oxidized to acetate and CO₂, and the electrons were quantitatively transferred to the working electrode. Growth yields of 29.0, 13.4 and 14.2 g cell material per mol were calculated, respectively. The high cell yield obtained during propionate oxidation cannot be explained solely by substrate level phosphorylation indicating that additional energy was conserved via electron transport phosphorylation. Furthermore, this result indicated complete reversibility of the methyl-malonyl-CoA pathway in propionic acid bacteria.     

Immunological studies of ferredoxin-nitrite reductases and ferredoxin-glutamate synthases from photosynthetic organisms

Polyclonal antiserum specific for ferredoxin-nitrite reductase (EC 1.7.7.1) from the green alga Chlamydomonas reinhardii recognized the nitrite reductase from other green algae, but did not cross-react with the corresponding enzyme from different cyanobacteria or higher plant leaves. An analogous situation was also found for ferredoxin-glutamate synthase (EC 1.4.7.1), using its specific antiserum. Besides, the antibodies raised against C. reinhardii ferredoxin-glutamate synthase were able to inactivate the ferredoxin-dependent activity of nitrite reductase from green algae.These results suggest that there exist similar domains in ferredoxin-nitrite reductases and ferredoxin-glutamate synthases from green algae. In addition, both types of enzymes share common antigenic determinants, probably located at the ferredoxin-binding domain. In spite of their physicochemical resemblances, no apparent antigenic correlation exists between the corresponding enzymes from green algae and those from higher plant leaves or cyanobacteria.Abbreviations Fd ferredoxin - GOGAT glutamate synthase - MV⁺ reduced methyl viologen (radical cation) - NiR nitrite reductase - PMSF phenylmethylsulphonyl fluoride - SDS sodium dodecyl sulfate     

Production of novel pullulanases at high concentrations by two newly isolated thermophilic clostridia

Two thermophilic bacteria, which are capable of growing on starch at 60-70 degrees C under anaerobic conditions, were isolated from a sugar refinery in Uelzen and from Solar lake in Israel. On the basis of their physiological characteristics they were identified as Clostridium thermohydrosulfuricum Uel 1 and C. thermohydrosulfuricum Sol 1, respectively. The product pattern of glucose polymer hydrolysis showed that both strains secreted enzymes that possess amylolytic and pullulytic activities. The major product formed was maltose. In addition, alpha-glucosidase activity could be detected in the supernatants of Uel 1 strain. Compared to most anaerobes investigated these isolates secreted extremely high concentrations of pullulanases in batch culture. Up to 85% of the total enzyme synthesized was detected in the culture fluid. Unlike the pullulanases of type I, which can only attack the alpha-1,6-glycosidic linkages, the pullulanases of both clostridial strains were also capable of hydrolyzing alpha-1,4-linkages. The enzyme system of both bacteria was found to be highly thermoactive; optimal activity was detected at pH 5.0 and 85 degrees C. Even at 95 degrees C and without the addition of metal ions still 15% to 25% of enzymatic activity was detectable.     

Adenylyl cyclase in yeast: antibodies and mutations identify a regulatory domain

The adenylyl cyclase system of the yeast Saccharomyces cerevisiae contains the CYR1 polypeptide, responsible for catalyzing formation of cAMP from ATP, and two RAS polypeptides, responsible for stimulation of cAMP synthesis by guanine nucleotides. We have obtained rabbit antibodies that recognize the CYR1 protein. Antibodies were raised against synthetic oligopeptides and against a recombinant beta-galactosidase/CYR1 fusion protein. These antibodies have allowed the identification of the CYR1 gene product as a 205 kDa protein. Treatment with trypsin (2 micrograms/ml) reduced the size of the CYR1 protein from 205 to 155 kDa and produced an activated enzyme which no longer responded to guanine nucleotides. This result is consistent with a model in which adenylyl cyclase activity is regulated by an inhibitory domain near the amino-terminus of the CYR1 protein. This model is further supported by the finding that adenylyl cyclase activity is also markedly elevated and unresponsive to guanine nucleotides in mutant yeast strains that express only the carboxy-terminal half of the CYR1 protein. Treatment with high trypsin concentrations (greater than 10 micrograms/ml) caused release of adenylyl cyclase activity from the membrane. Comparison of immunoreactive CYR1 fragments released by trypsin and membrane bound genetically altered proteins suggests that the CYR1 protein is attached to the membrane via a separate trypsin sensitive anchoring protein rather than via a membrane anchoring domain.     

Voltammetric studies on the electrochemical behaviour of membrane-entrapped hemes

Summary The electrochemical behaviour of Fe(III)-protoporphyrin IX entrapped into a cellulose triacetate membrane has been investigated by cyclic voltammetry. The physical entrapment into a solid matrix does not modify the redox properties of the entrapped berries, which also act as efficient promoters in the electrochemistry of cytochromec. Such a system represents a promising example of a simple solid-state promoter, and stimulates further investigations in order to obtain more complex systems that may be of significance for basic and applied bioelectrochemistry.     

Leaf ultrastructure of C4 species possessing different Kranz anatomical types in the cyperaceae

The leaf ultrastructure of NADP-malic enzyme type C₄ species possessing different anatomical features in the Cyperaceae was examined: types were the Rhynchosporoid type, a normal Kranz type in which mesophyll cells are adjacent to Kranz cells, and Fimbristyloid and Chlorocyperoid types, unusual Kranz types in which nonchlorophyllous mestome sheath intervenes between the two types of green cells. They show structural characteristics basically similar to the NADP-malic enzyme group of C₄ grasses, that is, centrifugally located chloroplasts with reduced grana and no increase of mitochondrial frequency in the Kranz cells. However, the Kranz cell chloroplasts of the Fimbristyloid and Chlorocyperoid types exhibit convoluted thylakoid systems and a trend of extensive development of peripheral reticulum, although those of the Rhynchosporoid type do not possess such particular membrane systems. The suberized lamella, probably a barrier for CO₂ diffusion, is present in the Kranz cell walls of the Rhynchosporoid type and in the mestome sheath cell walls of the other two types, and tightly surrounds the Kranz cells (sheaths) that are the sites of the decarboxylation of C₄ acids. These ultrastructural features are discussed in relation to C₄ photosynthetic function.     

The ecology of a beech forest on Mt. Sanpoiwadake,Hakusan National Park,Japan II. The correlation of the subassociation within the Lindero membranaceae-fagetum crenatae association and environmental parameters

Detailed examination of a sample plot covered by the Lindero membranaceae-Fagetum crenatae association on Mt. Sanpoiwadake, Hakusan National Park, revealed a number of correlations between the distribution of subassociations and environmental factors. The subassociations on the south-facing slopes receive deep snow cover in winter with rapid melting in the spring. They occur on porous, freely draining soils, typical of the general range of brown forest soils. Conversely, on the north-eastern slopes there are widespread late-snow patches which delay leaf development and expansion and which provide an abundant water supply well into early summer. Under these conditions, bleached soil horizons have developed with iron pan formation, resulting in poor soil drainage, strongly correlated with quite different plant communities.