Peptide des β-Alanins als Ersatz für die freie Aminosäure bei pantothensäurebedürftigen Hefezellen und ihr Verhalten gegenüber dem β-Alanin-Antagonisten Asparagin

Zusammenfassung Pantothensäurebedürftige Hefezellen können ihren Bedarf an diesem Vitamin nicht allein aus -Alanin decken, sondern auch aus Benzoyl--Alanin, -Alanyl-d,l-Norleucin und -Alanyl-l-Histidin. Der Antagonist Asparagin hemmt die Verwertung dieser Peptide genauso wie diejenige der freien Aminosäure. Durch höhere Konzentrationen an -Alanin oder -Alanyl-d,l-Norleucin läßt sich die Hemmwirkung nicht allein kompensieren, es kommt sogar zu einer Förderung des Hefewachstums. Der Antagonist wird dann zum Synergisten.

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Summary The -alanine containing peptides benzoyl--alanine, -alanyl-d,l-norleucine and -alanyl-l-histidine can substitute for the amino acid -alanine in a pantothenic acid requiring yeast. Asparagine, an antagonist of -alanine, affects these peptides in a similar manner. In combination with an overdose of -alanine or -alanyl-d,l-norleucine, asparagine is no longer an antagonist but becomes a synergist.

     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

Evolution of a protein superfamily: Relationships between vertebrate lens crystallins and microorganism dormancy proteins

Summary A search of sequence databases shows that spherulin 3a, an encystment-specific protein ofPhysarum polycephalum, is probably structurally related to the - and -crystallins, vertebrate ocular lens proteins, and to Protein S, a sporulation-specific protein ofMyxococcus xanthus. The - and -crystallins have two similar domains thought to have arisen by two successive gene duplication and fusion events. Molecular modeling confirms that spherulin 3a has all the characteristics required to adopt the tertiary structure of a single -crystallin domain. The structure of spherulin 3a thus illustrates an earlier stage in the evolution of this protein superfamily. The relationship of - and -crystallins to spherulin 3a and Protein S suggests that the lens proteins were derived from an ancestor with a role in stressresponse, perhaps a response to osmotic stress.     

Moraxellae: Identification

According to published reports, all strains ofMoraxella osloensis and Moraxella (Oligella) urethralis grew on a mineral-base medium supplemented with 0.015M (0.123%) sodium acetate, but not all alkalinized a mineral-base medium supplemented with 0.008% bromthymol blue and 0.2% sodium acetate. This seeming discrepancy was examined. The growth of most strains ofM. urethralis was inhibited by 0.008% bromthymol blue, and some by 0.2% anhydrous sodium acetate. All (42) strains ofM. osloensis andM. urethralis alkalinized a mineral-base medium supplemented with 0.1% sodium acetate trihydrate and 0.001% bromthymol blue or phenol red.     

Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae

The presence of active mitochondria and oxidative metabolism is shown to be essential to maintain low inhibition levels by ethanol of the growth rate (), fermentation rate (v) or respiration rate () of Saccharomyces cerevisiae wild type strain S288C. Cells which have respiratory metabolism show K _i (ethanol inhibition constant) values for , v and , higher (K _i>1 M) than those of petite mutants or grande strains grown in anaerobiosis (K _i=0.7 M). In addition, the relationship between or v and ethanol concentration is linear in cells with respiratory metabolism and exponential in cells lacking respiration. When functional mitochondria are transferred to petite mutants, the resulting strain shows K _i values similar to those of the grande strain and the inhibition of and v by increasing ethanol concentrations becomes linear.     

Comparative study on the chloroplast,mitochondrial and nuclear genome differentiation in two cultivated rice species,Oryza sativa and O. glaberrima,by RFLP analyses

Summary Restriction fragment length polymorphisms of chloroplast (ct), mitochondrial (mt) and nuclear DNA were investigated using eight cultivars of Oryza sativa and two cultivars of O. glaberrima. Relative variability in the nuclear and cytoplasmic genomes was estimated by a common measure, genetic distance. Based on the average genetic distances among ten cultivars for each genome, the evolutionary variabilities of the mitochondrial and nuclear genomes were found to be almost the same, whereas the variability of the chloroplast genome was less than half that of the other two genomes. Cluster analyses on ct and mt DNA variations revealed that chloroplast and mitochondrial genomes were conservative within a taxon and that their differentiations were well-paralleled with respect to each other. For nuclear DNA variation, an array of different degrees of differentiation was observed in O. sativa, in contrast with little variation in O. glaberrima. As a whole, differentiation between O. sativa and O. glaberrima was clearly observed in all three genomes. In O. sativa, no notable difference was found between the cultivars Japonica and Javanica, whereas a large differentiation was noticed between Japonica (including Javanica) and Indica. In all three genomes, the average genetic distances within Indica were much larger than those within Japonica (including Javanica), and almost similar between Japonica (including Javanica) and Indica. These facts indicate that differentiation in O. sativa was due mainly to Indica.     

Endogenous abscisic acid and 2-trans-abscisic acid in alternate bearing ‘Wilking’ mandarin trees

The relationship of abscisic acid (ABA) and 2-trans-abscisic acid (t-ABA) to alternate bearing has been examined in Wilking mandarin (Citrus reticulata Blanco) trees. Leaves, stems and buds of trees loaded with fruit (on trees) had 4.3, 6.0 and 2.2 fold higher ABA levels than the corresponding organs from off trees. Leaves had higher ABA levels than stems and buds in both on and off trees. t-ABA was non-detectable in Wilking leaf, stem and bud tissue. Amounts of t-ABA not exceeding 40% of the ABA content, were found in Shamouti and Valencia orange buds and in Wilking fruit peel.The elevated levels of ABA in on tree organs may reflect a stress imposed by the fruit overload.     

Substrate specificity and distribution of acid β-galactosidase activities in seizure-susceptible and non-susceptible strains of mice

The properties and distribution of -galactosidase were studied in the mouse brain using the artificial substrate methylumbelliferyl--galactoside. Enzyme activities were compared between an audiogenic seizure-susceptible mouse strain (DBA/2) and three non-susceptible strains of mice (BALB/c, C3H/He and Swiss A2G). At all ages, DBA/2 mice have significantly lower -galactosidase activity compared with the three other mouse strains: this is attributed to the different alleles present at the Bgs locus. The low activity of -galactosidase is also evident when the natural substrate GMI-ganglioside is hydrolyzed. In contrast to this low GMI-ganglioside--galactosidase activity, there is no difference in the activity of the second form of acid -galactosidase, galactosylceramidase, in DBA/2 mice at 7 and 14 days. However, at 21 and 28 days the activity is significantly lower in DBA/2 mice compared with the other strains of mice. These results on -galactosidase activity in the brain of seizure-susceptible and non-susceptible mice are discussed in relation to published levels of GMI-ganglioside and galactosylceramide present in the developing mouse brain.Dedicated to Henry McIlwain.     

Holling's “hungry mantid” model for the invertebrate functional response considered as a Markov process. Part II: Negligible handling time

In this paper we analyse a stochastic model for invertebrate predation taking account of the predator's satiation. This model approximates Holling's hungry mantid model when handling time is negligible (see Part I). For this model we derive equations from which we can calculate the functional response and the variance of the total catch. Moreover we study a number of approximations which can be used to calculate these quantities in practical cases in a relatively simple manner.List of Notation a rate constant of digestion - b maximum of rate constant of prey encounter in the mantid - c satiation threshold for search - c satiation threshold for pursuit in the mantid - c _i (w^1/2(N- N)ⁱ) - expectation operator - f rate of change of satiation during search - F functional response: mean number of prey eaten per unit of time - g rate constant of prey capture - h probability generating function of N conditional on S = s times p - H probability generating function of N - m_i ¹ - n, N number of prey caught - p probability density of S - p_n simultaneous probability (density) of N and S - q probability of strike success - r dummy variable in generating function - s, S satiation - T _s search time - T _d digestion time - v asymptotic rate of increase of var v - V asymptotic rate of increase of var N - w weight of edible part of prey - W standard Wiener process - x prey density - z (N{S = s}-N)p - rate constant of prey escape time maximum pursuit time - (v{S = + w ^1/2}-v) - present time as a fraction of the time from the start to the end of the experiment - hazard rate of T _s - mean time between (downward) passages of S through c - v w_–1/2(N-) - edible prey biomass density - probability density of , number pi - parameter of Weibull distribution of T _s = (1/2acx(-g(c)))_1/2 - w_–1/2(S -) - satiation in the guzzler approximation: solution to d/dt = f() + g(), (0)=S(0). - biomass functional response: wF - total biomass catch in the guzzler approximation: solution to d/dt = g(), (0) = 0     

Das organische Gerüstwerk der Kalkschale des Schwanen-Eies

Zusammenfassung Kegel und Säulen der Schwanen-Eischale hinterlassen am Querschliff nach Entkalkung mit EDTA organisches (mucoproteides) Material als ein zusammenhängendes Gerüst, das sich mit Thionin metachromatisch färbt; ohne Demineralisierung oder wenigstens Anätzung bleibt Thionin an Schliffen und Bruchkanten der Schale wirkungslos. Das Lichtmikroskop zeigt an Schliffen nichts von dem organischen Material, es wurde während des Kristallwachstums fein zerteilt in Gitterlücken des Schalencalcits eingeschlossen. Es findet sich am stärksten angehäuft an den äueren und inneren Oberflächen der Kristall-individuen. In den Kegeln ist das Gerüst radial ausgebildet als die Loculi der Keile, und konzentrisch geschichtet, entsprechend den Lagen der Globularinklusionen, um deren jede herum Verdichtung der organischen Substanz statthat. In den inneren Säulen folgt das organische Gerüst dem Rhombenmuster; die äueren Säulen sind arm an organischer Substanz, hier verbleibt nach der Entkalkung eine dünne laterale Oberflächenschicht.

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Summary The cones and columns of the swans egg shell leave behind after decalcification with EDTA an organic (mucoproteid) material in form of a continuous frame work stainable metachromatically with thionine. Without demineralisation or at least etching, thionine proves ineffectual in ground sections or breaking edges of the shell. In ground sections the light microscope demonstrates nothing of the organic material: it was inclosed during the crystal growth in submicroscopical lattice gaps of the calcite individuals. The organic material is chiefly accumulated in the outer and inner surfaces of the crystals. In the cones the organic frame work is developed radially as the loculi of the wedges and concentrically layered corresponding with the globular inclusions, concentrated in the circumference of each. In the inner columns the organic material follows to the rhomb pattern. The outer columns after decalcification only leave behind a thin lateral organic sheath.

     

Forskolin Modulates Acetylcholine Receptor Gating by Interacting with the Small Extracellular Loop Between the M2 and M3 Transmembrane Domains

1. Forskolin acts as an allosteric modulator of muscle-type nicotinic acetylcholine receptors. Receptors from mouse muscle and Torpedo electroplax demonstrate differential sensitivity to inhibition by forskolin. Previous work from this laboratory suggested that the subunit is responsible for this differential sensitivity.2. We have used a series of mouse/Torpedo species-chimeric subunits to further define the site of forskolin interaction with the subunit. Analysis of the patterns of forskolin inhibition of receptors containing mouse/Torpedo chimeric subunits along with the mouse , , and subunits suggests that forskolin interacts with the small extracellular domain that links the M2 and M3 transmembrane domains (the M2–M3 linker).3. We suggest that the M2–M3 linker domain plays an important role in the transduction of ligand binding to the conformational changes that result in channel opening.     

Variability in the 11S globulin fraction of seed storage proteins ofHelianthus (Asteraceae)

The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (₁₁ and ₂₂). The subunit corresponding to the ₁₁ pair is present inH. petiolaris and in the three populations ofH. annuus studied. The _2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific _2a2 and ₄₄ pairs andH. annuus a specific ₃₃ pair. InH. argophyllus _{11 33} or ₄₄ are never observed but are replaced by ₁₃ and ₃₁ pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins.     

Purification of alpha-hemolysin from an overproducing E. coli strain

Summary The genetic determinant of the -hemolysin encoded by plasmid pHly152 has been cloned in both orientations in plasmid pBR322 giving rise to plasmids pSU157 and pSU158. E. coli strains carrying either of these recombinant Hly plasmids produced about 20 times more hemolysin activity than the parental plasmid pHly152, when grown in minimal medium supplemented with hemoglobin. Thus high hemolytic activity is not lethal to the cells, contrary to previous assumptions.-hemolysin was purified from culture supernatants of strain SU100 (pSU157) by ammonium sulfate precipitation and gel filtration in Sephacryl S-200 in the presence of 6 M urea. When purified -hemolysin preparations were subjected to electrophoretic analysis in denaturing conditions, a single 107 kdal polypeptide was observed. This probably corresponds to the -hemolysin protein, since an isogenic E. coli strain carrying plasmid pSU161, an Hly^- mutant derivative of pSU157, did not synthesize the 107 kdal polypeptide.     

Killing ofLegionella pneumophila by human serum and iron-binding agents

The inhibitory effect of serum on the growth and survival ofLegionella pneumophila Bloomington 2 was investigated. When incubated in the presence of 20%–50% normal human serum for 10 h, viability was decreased by >99%. Heat-inactivated or <40% normal serum supplemented with 50 M iron was not inhibitory. The addition of guinea pig complement to heat-inactivated serum resulted in killing of approximately 98% of the cells. Growth in buffered yeast extract broth was inhibited by the addition of ferric iron-binding compounds. Minimum bactericidal concentrations at 37°C were 10 M apotransferrin, 35 M 1,10-phenanthroline, and 50 M deferoxamine. Addition of iron chelators to normal serum did not accelerate killing. Egg yolk-passaged virulent strains and agar-grown avirulent strains exhibited similar serum sensitivity. Results of this study indicate that complement and serum transferrin are antagonistic to the growth ofLegionella in serum.     

β-Carotene producing mutants of Phaffia rhodozyma

Like other carotenoid-producing organisms, Phaffia rhodozyma, a red astaxanthin-producing yeast, is supposed to synthesize carotenoids by the following steps: formation of phytoene from geranylgeranyl pyrophosphate, dehydrogenation of phytoene to lycopene, cyclization of lycopene to -carotene and oxidation of the latter to astaxanthin. Mutagenic treatments generated in P. rhodozyma a wide diversity of colour variants ranging from white to dark red. The identification of the corresponding carotenoid compounds revealed the occurrence of -carotene-accumulating strains, phytoene-accumulating strains, and strains lacking any carotenoid compound. These classes of strains are likely to result from alterations in, respectively, the oxidation of -carotene, phytoene dehydrogenation and the phytoene synthetase step. Except for the cyclization of lycopene to -carotene, all the steps of carotenogenesis in P. rhodozyma are represented by the above mutants. Furthermore, astaxanthin-overproducing mutants were also selected; they are likely to be affected in some upstream step, and certainly before -carotene, as after an additional mutagenesis they generated oxidaseless strains that, in this case, overproduce -carotene. The latter strains appear very promising for biotechnological production of natural -carotene.     

DNA polymorphisms in the ψβ1- and β-globin gene regions in asian macaques

DNA polymorphisms in the ₁--globin gene region in nine Asian macaques(Macaca fuscata, M. mulatta, M. nemestrina, M. cyclopis, M. fascicularis, M. arctoides, M. radiata, M. maura, andM. assamensis) were examined using several restriction endonucleases and the human ₁, IVS2, and IVS2 probes. TheBamHI site 3 to the -globin gene was polymorphic inM. fuscata andM. mulatta, while the HincII site and the EcoRI site in the ₁-globin gene region was highly polymorphic inM. fuscata andM. mulatta, respectively. These polymorphic sites also seem to be present in other Asian macaques. The present study of the polymorphism at theBamHI site 3 to the -globin gene in Asian macaques supports, at the nuclear DNA level, the idea that thefascicularis group includingM. fuscata, M. mulatta, M. cyclopis, andM. fascicularis is different from other Asian macaque groups.This study was supported in part by the Cooperation Research Program of the Primate Research Institute, Kyoto University.     

Synthetic multifunctional proteins

Summary Several E. coli mutants were isolated which produce triple chimeras between one of the trp enzymes lac, repressor and -galactosidase. The mutants were isolated as TonB^- Lac⁺ derivatives of a phenotypically Lac^- TrpR^- strain carrying a lac I ⁺-Z⁺ fusion on a 80dlac phage. The phage is integrated into the chromosome in such a way that the lac and the trp genes are transcribed in the same direction. Of a total of 58 candidates 2 TrpA^- and 3 Trp^- strains produce triple chimeras. The chimeras from the two TrpA^- strains were further examined. They consist of tryptophan synthetase -subunit, lac repressor and -galactosidase. In crude extracts of these strains the tryptophan synthetase -subunit part can be identified by its ability to aggregate with the -subunit since some of the -subunit activity can be precipitated with antiserum against -galactosidase. Furthermore -galactosidase precipitates with antiserum against tryptophan synthetase -subunit. The lac repressor part is able to bind IPTG, but not lac operator DNA in vitro. The -galactosidase part is as unaffected as in the original lac repressor--galactosidase chimera. The molecular weigths of both chimeras are 175,000 when determined by SDS gel electrophoresis. The chimeras are partially degraded giving rise to fragments of distinct molecular weights.     

Vertebrate-like βγ-crystallins in the ocular lenses of a copepod

The diverse crystallins are water-soluble proteins that are responsible for the optical properties of cellular lenses of animal eyes. While all vertebrate lenses contain physiological stress-related - and -crystallins, some also contain taxon-specific, often enzyme-related crystallins. To date, the - and -crystallins have been found only in vertebrate lenses. Here we report lenses from an invertebrate, the pontellid copepod Anomalocera ornata, accumulate -crystallin family members as judged by immunocytochemistry, western immunoblotting and microsequencing. Our data provide the first example of -crystallin members in an invertebrate lens, establishing that the use of this protein family as lens crystallins is not confined to vertebrates.