Purification and properties of gentisate 1,2-dioxygenase from Moraxella osloensis.

Gentisate:oxygen 1,2-oxidoreductase (decyclizing) (EC 1.13.11.4; gentisate 1,2-dioxygenase) from Moraxella osloensis was purified to homogeneity as shown by polyacrylamide gel electrophoresis. The enzyme has a molecular weight of about 154,000 and gives rise to subunits of molecular weight 40,000 in the presence of sodium dodecyl sulfate. Gentisate 1,2-dioxygenase showed broad substrate specificity and attacked a range of halogen- and alkyl-substituted gentisic acids. Maleylpyruvate, the product formed from gentisate, was degraded by cell extracts supplemented with reduced glutathione, but substituted maleylpyruvates were not attacked under these conditions.     

Patterns of hemolymph osmoregulation in three desert arthropods

Summary Hemolymph osmoregulation was examined in three species of desert arthropods: a tenebrionid beetle,Eleodes hispilabris, a vejovid scorpion,Paruroctonus aquilonalis, and a spirostreptid millipede,Orthoporus ornatus. During desiccation, beetles regulated hemolymph osmolality and scorpions tolerated increasing osmolality. Millipedes displayed both osmotic regulation and tolerance patterns depending on sex and on duration of desiccation. Rehydration after desiccation depressed blood osmolality of male scorpions and beetles below levels found for freshly collected specimens. This was not the case in millipedes. Seasonal osmolality changes were studied and recorded among field-collected scorpions and beetles. Patterns of regulation and tolerance of hemolymph osmolality appear to vary among different kinds of desert arthropods.     

A numerical study of the common narrow-leaved taxa of chenopodium occurring in the Western United States

A numerical taxonomic study was conducted on the common narrow-leaved members of the genusChenopodium that occur in the western United States. As many as six and as few as two species have been recognized. Thirty-five individual popu lations were used as OTU’s, and 35 characters were considered. The data were subjected to cluster (both the weighted-pair group method and the within-group dispersion method) and principal component analysis. The results of both analyses are in general agreement and suggest phenetic relationships that differ from current interpretations in the literature. It is concluded thatC. desiccatum var.leptophylloides is much more closely related phenetically toC. atrovirens (and may not be distinct from it) than it is toC. desiccatum var.desiccatum. Chenopodium hians andC. leptophyllum appear to be phenetically distinct despite the fact that they have sometimes been viewed as conspecific.Cheno podium subglabrum is not very similar toC. leptophyllum, and should probably be treated as a separate species rather than as a variety of the latter. Popu lations referable toC. incognitum form a rather loose cluster that appears some what intermediate between populations ofC. desiccatum var.leptophylloides,C. atrovirens, C. hians, andC. leptophyllum.     

Water relations and desiccation-avoiding behavior in the vinegaroon Mastigoproctus giganteus (Arachnida: Uropygi)

The vinegaroon or whip scorpion Mastigoproctus giganteus gains water by drinking from a moist substrate and from its prey. These are found by the sensitive antenniform front legs. Large specimens readily lose water to the air at 26° at humidities up to 95% R.H. As temperatures increase they lose water in proportion to the saturation deficit of the atmosphere, suggesting the absence of an epicuticular wax layer.Desiccated vinegaroons respond positively to moist air in a humidity gradient, but show no preference between darkness and light. Substrate moisture does not affect the lack of such a preference by normal animals.Burrowing is characteristic of dehydrated individuals. By using their sensitive front legs these first detect nonhorizontal surfaces and, provided the substrate is moist, dig burrows against — and in nature most likely beneath — such surfaces so that further water loss is presumably avoided.

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Résumé Mastigoproctus giganteus se rencontre dans les régions arides du Sud-Ouest des U.S.A.; rare pendant la saison sèche, il appraît après les pluies. C'est un prédateur nocturne. principalement d'insectes. Le présent travail tente de préciser comment cet Arachnide réagit aux conditions d'aridité des régions désertiques.De grands exemplaires vivants de M. giganteus montrent un taux de perte en eau relativement élevé pour une température de 26° avec une humidité relative pouvant aller jusqu'à 95%. Pour ces grands spécimens la transpiration à travers la cuticule s'accroît de façon approximativement linéaire pour les températures supérieures, ce qui indique l'absence probable d'une couche cireuse épicuticulaire fonctionnelle. Des animaux hydratés ne répondent pas à un gradient d'humidité, mais après avoir été soumis à des conditions de dessication les individus de petite taille montrent une préférence marquée pour l'air le plus humide. Ils ne manifestent aucune préférence entre la lumière (1,500 lux) ou l'obscurité lorsqu'ils sont placés sur sable sec ou humide, même après avoir une dessication partielle.La faible aptitude physiologique de ces animaux pour réduire leurs pertes en eau semble compensée par un sens thigmotactique hautement développé qui leur permet d'une part, de capturer aisément leurs proies, d'autre part de détecter des surfaces non-horizontales contre lesquelles ils creusent alors si le substrat est humide. Ce comportement doit jouer un rôle important en évitant à l'animal les conditions de dessication.

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This work was carried out during tenure of a National Science Foundation Senior Visiting Scientist Fellowship, for which appreciative acknowledgment is hereby given.     

Relation entre réaction géotropique et évolution du statenchyme dans la racine d'Asperge

Relationship between the Geotropic Response and the Evolution of the Statenchyma in Roots of Asparagus officinalis. The evolution of the statenchyma in roots of Asparagus of ficinalis seedlings, grown in obscurity, was followed during the first 17 days. After 7 days of etiolation, a decrease of both the average diameter of the amyloplasts and the average number of these organelles was observed in the central root cap cells. If the seedlings were illuminated (with white light) from the 7th day, the average number of statoliths increased rapidly in the statocytes. The volume of these organelles undergoes the same variation in etiolated and in illuminated plants. The initial rate of curvature (V_i) of the roots (stimulated in a horizontal position) and the volume of amyloplasts (V_ac) in their caps were analysed as a function of the time of germination in obscurity (from the 8th to the 17th day). It was found that V_i increased as a linear function of the logarithm of V_ac, which confirms that the weight of the amyloplasts of the statocytes may play a role in the geotropic stimulation of the roots.     

A new model for cellulose architecture in some plant cell walls

Summary A new model of rotating fibre components (helicoidal model) is proposed to explain the architecture of some plant cell walls. On the basis of tilting observations under the electron microscope, we establish the validity of this model for the cell wall ofChara vulgaris oospores. We suggest that this model explains the architecture seen in a number of published micrographs from a variety of different plant cell walls. Helicoidal architecture is shown to be distinct from the previously established crossed polylamellate architecture. The diagnostic features of helicoidal architecture are given. Morphogenesis of plant cell walls is discussed, with particular reference to self assembly in cholesteric liquid crystals.     

Platelet contractile proteins: separation and characterization of the actin and myosin-like components.

Solution of thrombosthenin, the contractile protein complex isolated from pig platelets, have been studied by analytical ultracentrifugation and zone sedimentation in sucrose density gradients. Freshly prepared thrombosthenin in 0.6 M KCl shows a prominent peak in the ultracentrifuge with S degrees 20w about 5.5 and higher molecular weight aggregates (greater than 100S) sedimenting quickly to the bottom of the cell. Short term storage of high ionic strength solutions of thrombosthenin induces actomyosin-like gel formation and these gels dissociate with ATP and Mg2+ ions into two components of S degrees 20w 8.0 and S degrees 20w50. The supernatant, after actomyosin gel removal, contains only the S degrees 20w5.5 protein. From results of Ca2+ ATPase activity measurements and SDS polyacrylamide gel electrophoretic mobilities of dissociated thrombosthenin separated into fractions in sucrose density gradients, it is concluded that the S degrees20w5.5 protein species is the myosin-like protein of thrombosthenin. The S degrees 20w8.0 protein is not fibrinogen but also has myosin-like properties and is believed to be myosin dimer. Species of higher S values seen in the presence of ATP and Mg2+ in the analytical ultracentrifuge and located in the higher density zones of the sucrose gradients all gave in SDS polyacrylamide gel electrophoresis a single band of molecular weight 46-47,000 daltons. These subunit proteins appear to be derived from a range of polymeric variants of the F-actin-like protein of the contractile complex. All these higher density F-actin-like proteins readily form superprecipitates and display syneresis when combined with rabbit skeletal muscle myosin or platelet myosin. They are also all capable of conferring upon these two myosins a Mg2+ activated ATPase activity. It is suggested that in thrombosthenin solutions a myosin monomer-dimer equilibrium state exists which can be directionally influenced by a number of factors. The coexistence in the solution of F-actin and Mg2+ ATP, for example, increases the propensity of the myosin-like protein to form the higher molecular weight aggregate. Such aggregation may be the initiating mechanism for the intracellular organization of the thick filaments of the actomyosin complex, preparatory to a contractile event.     

Pseudomonas putida Tryptophan Synthetase

The two protein components of Pseudomonas putida tryptophan synthetase have been purified to homogeneity. Although there is general similarity between the Pseudomonas enzyme and that of the enteric bacteria, many differences were found. Components from Escherichia coli and P. putida do not stimulate each other enzymatically, and the enzymes differ in their response to monovalent cations. Serine deamination occurs best with the intact enzyme of P. putida, not with the beta(2) subunit alone as in E. coli. The amino acid compositions of the alpha subunits differ appreciably. These findings extend earlier studies showing differences between enteric organisms and pseudomonads in the regulation and genetic organization of the enzymes of the tryptophan pathway.     

Extracellular Enzyme Activities during Lignocellulose Degradation by Streptomyces spp.: A Comparative Study of Wild-Type and Genetically Manipulated Strains

The wild-type ligninolytic actinomycete Streptomyces viridosporus T7A and two genetically manipulated strains with enhanced abilities to produce a water-soluble lignin degradation intermediate, an acid-precipitable polymeric lignin (APPL), were grown on lignocellulose in solid-state fermentation cultures. Culture filtrates were periodically collected, analyzed for APPL, and assayed for extracellular lignocellulose-catabolizing enzyme activities. Isoenzymes were analyzed by polyacrylamide gel electrophoresis and activity staining on the gels. Two APPL-overproducing strains, UV irradiation mutant T7A-81 and protoplast fusion recombinant SR-10, had higher and longer persisting peroxidase, esterase, and endoglucanase activities than did the wild-type strain T7A. Results implicated one or more of these enzymes in lignin solubilization. Only mutant T7A-81 had higher xylanase activity than the wild type. The peroxidase was induced by both lignocellulose and APPL. This extracellular enzyme has some similarities to previously described ligninases in fungi. This is the first report of such an enzyme in Streptomyces spp. Four peroxidase isozymes were present, and all catalyzed the oxidation of 3,4-dihydroxyphenylalanine, while one also catalyzed hydrogen peroxide-dependent oxidation of homoprotocatechuic acid and caffeic acid. Three constitutive esterase isozymes were produced which differed in substrate specificity toward α-naphthyl acetate and α-naphthyl butyrate. Three endoglucanase bands, which also exhibited a low level of xylanase activity, were identified on polyacrylamide gels as was one xylanase-specific band. There were no major differences in the isoenzymes produced by the different strains. The probable role of each enzyme in lignocellulose degradation is discussed.     

Viscoelastic relaxation of bilayer lipid membranes. Frequency-dependent tension and membrane viscosity.

Photon correlation spectroscopy has been used to study capillary waves on black lipid membranes of glycerol monooleate at temperatures above the lipid transition. For the first time the tension and viscosity of solvent-free bilayers have been observed to display a frequency dependence. The variations of both parameters can be accounted for by a Maxwell viscoelastic fluid model having a relaxation time of 37 microseconds. The equilibrium (omega = 0) tension is compatible with literature values. The present results do not suffice to precisely define the specific molecular processes involved, but relaxation times similar to the present are associated with certain phenomena in phospholipid vesicles. Bilayers containing hydrocarbon solvent do not show such relaxation, presumably due to their weaker intermolecular interactions.