TY - JOUR A1 - Wallschläger, Hans-Dieter A1 - Beier, Wolfgang A1 - Burkart, Michael A1 - Mrzljak, Jadranka A1 - Oehlschläger, Susanne A1 - Wanner, Manfred T1 - Ökologische Datenerfassung für Naturschutzbewertung und Monitoring im Offenland Y1 - 2004 SN - 3-540-22449-1 ER - TY - JOUR A1 - Seitz, Birgit A1 - Ristow, Michael A1 - Klemm, Gunther A1 - Rätzel, Stefan A1 - Schulze, Gerhart A1 - Hoffmann, Maik T1 - Zur Verbreitung der Wildrosen und verwilderten Kulturrosen in Berlin und Brandenburg Y1 - 2004 SN - 0724-3111 - ER - TY - JOUR A1 - Albrecht, Tanja A1 - Haebel, Sophie A1 - Koch, Anke A1 - Krause, Ulrike A1 - Eckermann, Nora A1 - Steup, Martin T1 - Yeast glycogenin (Glg2p) produced in Escherichia coli is simultaneously glucosylated at two vicinal tyrosin residues but results in a reduced bacterial glycogen accumulation N2 - Saccharomyces cerevisiae possesses two glycogenin isoforms (designated as Glg1p and Glg2p) that both contain a conserved tyrosine residue, Tyr232. However, Glg2p possesses an additional tyrosine residue, Tyr230 and therefore two potential autoglucosylation sites. Glucosylation of Glg2p was studied using both matrix-assisted laser desorption ionization and electrospray quadrupole time of flight mass spectrometry. Glg2p, carrying a C-terminal (His(6)) tag, was produced in Escherichia coli and purified. By tryptic digestion and reversed phase chromatography a peptide (residues 219-246 of the complete Glg2p sequence) was isolated that contained 4-25 glucosyl residues. Following incubation of Glg2p with UDPglucose, more than 36 glucosyl residues were covalently bound to this peptide. Using a combination of cyanogen bromide cleavage of the protein backbone, enzymatic hydrolysis of glycosidic bonds and reversed phase chromatography, mono- and diglucosylated peptides having the sequence PNYGYQSSPAM were generated. MS/MS spectra revealed that glucosyl residues were attached to both Tyr232 and Tyr230 within the same peptide. The formation of the highly glucosylated eukaryotic Glg2p did not favour the bacterial glycogen accumulation. Under various experimental conditions Glg2p-producing cells accumulated approximately 30% less glycogen than a control transformed with a Glg2p lacking plasmid. The size distribution of the glycogen and extractable activities of several glycogen-related enzymes were essentially unchanged. As revealed by high performance anion exchange chromatography, the intracellular maltooligosaccharide pattern of the bacterial cells expressing the functional eukaryotic transgene was significantly altered. Thus, the eukaryotic glycogenin appears to be incompatible with the bacterial initiation of glycogen biosynthesis Y1 - 2004 ER - TY - JOUR A1 - Pacholsky, Dirk A1 - Vakeel, Padmanabhan A1 - Himmel, Mirko A1 - Lowe, T. A1 - Stradal, T. A1 - Rottner, K. A1 - Fürst, Dieter Oswald A1 - vanderVen, Peter F. M. T1 - Xin repeats define a novel actin-binding motif N2 - Xin is a protein that is expressed during early developmental stages of cardiac and skeletal muscles. Immunolocalization studies indicated a peripheral localization in embryonic mouse heart, where Xin localizes with beta- catenin and N-cadherin. In adult tissues, Xin is found primarily in the intercalated discs of cardiomyocytes and the myotendinous junctions of skeletal muscle cells, both specialized attachment sites of the myofibrillar ends to the sarcolemma. A large part of the Xin protein consists of unique 16 amino acid repeats with unknown function. We have investigated the characteristics of the Xin repeats by transfection experiments and actin-binding assays and ascertained that, upon expression in cultured cells, these repeats bind to and stabilize the actin-based cytoskeleton. In vitro co- sedimentation assays with skeletal muscle actin indicated that they not only directly bind actin filaments, but also have the capability of arranging microfilaments into networks that sediment upon low-speed centrifugation. Very similar repeats were also found in Xin-repeat protein 2' (XIRP2), a novel protein that seems to be expressed mainly in striated muscles. Human XIRP2 contains 28 Xin repeats with properties identical to those of Xin. We conclude that the Xin repeats define a novel, repetitive actin-binding motif present in at least two different muscle proteins. These Xin- repeat proteins therefore constitute the first two members of a novel family of actin-binding proteins Y1 - 2004 SN - 0021-9533 ER - TY - JOUR A1 - Kummer, Volker T1 - Vom Seidenbau in Krausnick Y1 - 2004 ER - TY - JOUR A1 - Boese, Stefan H. A1 - Gray, Michael A. A1 - Simmons, N. L. T1 - Volume and non-volume activated anion conductances and their interactions in the renal IMCD Y1 - 2004 SN - 0-387- 23299-0 ER - TY - JOUR A1 - Weithoff, Guntram T1 - Vertical niche separation of two consumers (Rotatoria) in an extreme habitat N2 - Herbivore populations are commonly restricted by resource limitation, by predation or a combination of the two. Food supplement experiments are suitable for investigating the extent of food limitation at any given time. The main part of this study was performed in an extremely acidic lake (pH 2.7) where the food web consists of only a few components and potential food sources for herbivores are restricted to two flagellates. Life table experiments proved that Chlamydomonas was a suitable food source whereas Ochromonas was an unsuitable food source. The two flagellates and the two rotifers exhibit a pronounced vertical distribution pattern. In this study, a series of food supplement experiments were performed in order to: (1) quantify and compare potential resource limitation of two primary consumers (Cephalodella hoodi and Elosa worallii, Rotatoria) over time, (2) compare their response at different temperatures, (3) evaluate the effect of having an unsuitable food source alongside a valuable one, (4) estimate the effect of predation on rotifers by Heliozoa, and (5) compare the results with those from other acidic lakes. Additionally, the spatio- temporal population dynamics of both species were observed. The field data confirmed a vertical separation of the two species with E. worallii dominating in the upper water layers, and C. hoodi in the deeper, cooler water layers. The results from the food supplement experiments in which Chlamydomonas served as the supplemented suitable food source showed that the two rotifers were food limited in the epilimnion throughout the season to different extents, with Cephalodella being more severely food limited than Elosa. The experiments at different temperatures provided evidence that Elosa had a higher optimum temperature for growth than Cephalodella. When the unsuitable food algae Ochromonas was added alongside the suitable food source Chlamydomonas, C. hoodi was unaffected but E. worallii was negatively affected. Predation of Heliozoa on rotifers was observed but the total effect on the rotifer dynamics is probably low. The comparison with other lakes showed that resource limitation also occurred in one other lake, although to a lesser extent. Overall, the vertical separation of the two rotifers could be explained by both their differential extent of resource limitation and differential response to temperature. Y1 - 2004 ER - TY - JOUR A1 - Hilson, Pierre A1 - Allemeersch, Joke A1 - Altmann, Thomas A1 - Aubourg, Sebastien A1 - Avon, Alexandra A1 - Beynon, Jim A1 - Bhalerao, Rishikesh P. A1 - Bitton, Frederique A1 - Caboche, Michel A1 - Cannoot, Bernard A1 - Chardakov, Vasil A1 - Cognet-Holliger, Cecile A1 - Colot, Vincent A1 - Crowe, Mark A1 - Darimont, Caroline A1 - Durinck, Steffen A1 - Eickhoff, Holger A1 - deLongevialle, Andeol Falcon A1 - Farmer, Edward E. A1 - Grant, Murray A1 - Kuiper, Martin T. R. A1 - Lehrach, Hans A1 - Leon, Celine A1 - Leyva, Antonio A1 - Lundeberg, Joakim A1 - Lurin, Claire A1 - Moreau, Yves T1 - Versatile gene-specific sequence tags for arabidopsis functional genomics : transcript profiling and reserve genetics applications N2 - Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics Y1 - 2004 ER - TY - JOUR A1 - Sachinidis, A. A1 - Wartenberg, Maria A1 - Sauer, Heinrich A1 - Hescheler, Jürgen T1 - Transcription factors, growth factors and signal cascades capable of priming morphogenesis of heart Y1 - 2004 SN - 1-588- 29113-8 ER - TY - JOUR A1 - Witt, Isabell A1 - Zanor, Maria Ines A1 - Müller-Röber, Bernd T1 - Transcription factor function search : how do individual factors regulate agronomical important processes in plants? (Subproject A) Y1 - 2004 SN - 3-00-011587-0 ER - TY - JOUR A1 - Krylov, Andrey V. A1 - Beissenhirtz, Moritz Karl A1 - Adamzig, Holger A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Thick-film electrodes for measurement of superoxide and hydrogen peroxide based on direct protein-electrode contacts N2 - Cytochrome c was immobilized on screen-printed thick-film gold electrodes by a self-assembly approach using mixed monolayers of mercaptoundecanoic acid and mercaptoundecanol. Cyclic voltammetry revealed quasi-reversible electrochemical behavior of the covalently fixed protein with a formal potential of +10 mV vs. Ag/AgCl. Polarized at +150 mV vs. Ag/AgCl the electrode was found to be sensitive to superoxide radicals in the range 300-1200 nmol L-1. Compared with metal needle electrodes sensitivity and reproducibility could be improved and combined with the easiness of preparation. This allows the fabrication of disposable sensors for nanomolar superoxide concentrations. By changing the electrode potential the sensor can be switched from response to superoxide radicals to hydrogen peroxide-another reactive oxygen species. H2O2 sensitivity can be provided in the range 10-1000 mumol L-1 which makes the electrode suitable for oxidative stress studies Y1 - 2004 ER - TY - JOUR A1 - Hanisch, Uwe-Karsten A1 - Van Rossum, Denise A1 - Xie, Yiheng A1 - Misselwitz, Rolf A1 - Auriola, Seppo A1 - Goldstein, Gundars A1 - Koistinaho, Jari A1 - Kettemann, Helmut A1 - Möller, Thomas A1 - Gast, Klaus T1 - The microglia-activating potential of thrombin : the protease is not involved in the induction of proinflammatory cytokines and chemokines N2 - The serine protease thrombin is known as a blood coagulation factor. Through limited cleavage of proteinase- activated receptors it can also control growth and functions in various cell types, including neurons, astrocytes, and microglia ( brain macrophages). A number of previous studies indicated that thrombin induces the release of proinflammatory cytokines and chemokines from microglial cells, suggesting another important role for the protease beyond hemostasis. In the present report, we provide evidence that this effect is not mediated by any proteolytic or non- proteolytic mechanism involving thrombin proper. Inhibition of the enzymatic thrombin activity did not affect the microglial release response. Instead the cyto-/chemokine-inducing activity solely resided in a high molecular weight protein fraction that could be isolated in trace amounts even from apparently homogenous alpha- and gamma-thrombin preparations. High molecular weight material contained thrombin-derived peptides as revealed by mass spectrometry but was devoid of thrombin-like enzymatic activity. Separated from the high molecular weight fraction by fast protein liquid chromatography, enzymatically intact alpha- and gamma-thrombin failed to trigger any release. Our findings may force a revision of the notion that thrombin itself is a direct proinflammatory release signal for microglia. In addition, they could be relevant for the study of other cellular activities and their assignment to this protease Y1 - 2004 ER - TY - JOUR A1 - Hanisch, Uwe-Karsten A1 - van Rossum, D. A1 - Gast, Klaus A1 - Misselwitz, Rolf A1 - Goldstein, Gundars A1 - Koistinaho, Jari A1 - Möller, Thomas T1 - The microglia-activating potential of thrombin : is the protease able to induce cyto- and chemokines? Y1 - 2004 ER - TY - JOUR A1 - Scheffler, Christiane A1 - Ketelhut, Kerstin A1 - Mohasseb, Iman A1 - Ketelhut, Reinhard G. T1 - The influence of an exercise program on body composition, motor and cardiovascular parameters in pre-school children : a longitudinal study Y1 - 2004 SN - 88-87814-25-2 ER - TY - JOUR A1 - Fettke, Jörg A1 - Eckermann, Nora A1 - Poeste, Simon A1 - Steup, Martin T1 - The glycan substrate of the cytosolic (Pho 2) phosphorylase isozyme from Pisum sativum L. : identification, linkage analysis and subcellular localization N2 - The subcellular distribution of starch-related enzymes and the phenotype of Arabidopsis mutants defective in starch degradation suggest that the plastidial starch turnover is linked to a cytosolic glycan metabolism. In this communication, a soluble heteroglycan (SHG) from leaves of Pisum sativum L. has been studied. Major constituents of the SHG are galactose, arabinose and glucose. For subcellular location, the SHG was prepared from isolated protoplasts and chloroplasts. On a chlorophyll basis, protoplasts and chloroplasts yielded approximately 70% and less than 5%, respectively, of the amount of the leaf-derived SHG preparation. Thus, most of SHG resides inside the cell but outside the chloroplast. SHG is soluble and not membrane-associated. Using membrane filtration, the SHG was separated into a <10 kDa and a >10 kDa fraction. The latter was resolved into two subfractions (I and II) by field-flow fractionation. In the protoplast-derived >10 kDa SHG preparation the subfraction I was by far the most dominant compound. beta-Glucosyl Yariv reagent was reactive with subfraction II, but not with subfraction I. In in vitro assays the latter acted as glucosyl acceptor for the cytosolic (Pho 2) phosphorylase but not for rabbit muscle phosphorylase. Glycosidic linkage analyses of subfractions I and II and of the Yariv reagent reactive glycans revealed that all three glycans contain a high percentage of arabinogalactan-like linkages. However, SHG possesses a higher content of minor compounds, namely glucosyl, mannosyl, rhamnosyl and fucosyl residues. Based on glycosyl residues and glycosidic linkages, subfraction I possesses a more complex structure than subfraction II Y1 - 2004 SN - 0960-7412 ER - TY - JOUR A1 - Marg, S. A1 - Walz, Bernd A1 - Blenau, Wolfgang T1 - The effects of dopamine receptor agonists and antagonists on the secretory rate of cockroach (Periplaneta americana) salivary glands N2 - The acinar salivary glands of the cockroach, Periplaneta americana, are innervated by dopaminergic and serotonergic nerve fibers. Serotonin stimulates the secretion of protein-rich saliva, whereas dopamine causes the production of protein-free saliva. This suggests that dopamine acts selectively on ion-transporting peripheral cells within the acini and the duct cells, and that serotonin acts on the protein-producing central cells of the acini. We have investigated the pharmacology of the dopamine-induced secretory activity of the salivary gland of Periplaneta americana by testing several dopamine receptor agonists and antagonists. The effects of dopamine can be mimicked by the non-selective dopamine receptor agonist 6,7-ADTN and, less effectively, by the vertebrate D1 receptor-selective agonist chloro-APB. The vertebrate D1 receptor-selective agonist SKF 38393 and vertebrate D2 receptor-selective agonist R(-)- TNPA were ineffective. R(+)-Lisuride induces a secretory response with a slower onset and a lower maximal response compared with dopamine-induced secretion. However, lisuride-stimulated glands continue secreting saliva, even after lisuride-washout. Dopamine-induced secretions can be blocked by the vertebrate dopamine receptor antagonists cis(Z)- flupenthixol, chlorpromazine, and S(+)-butaclamol. Our pharmacological data do not unequivocally indicate whether the dopamine receptors on the Periplaneta salivary glands belong to the D1 or D2 subfamily of dopamine receptors, but we can confirm that the pharmacology of invertebrate dopamine receptors is remarkably different from that of their vertebrate counterparts. (C) 2004 Elsevier Ltd. All rights reserved Y1 - 2004 SN - 0022-1910 ER - TY - JOUR A1 - Sauer, Heinrich A1 - Wartenberg, Maria A1 - Sachinidis, A. A1 - Hescheler, Jürgen T1 - The development of the cardiovascular system in embryoid bodies deriverd from embryonic stem cells Y1 - 2004 SN - 1-588- 29113-8 ER - TY - JOUR A1 - Klein, Florian A1 - Feldhahn, Niklas A1 - Harder, S. A1 - Wang, Hui A1 - Wartenberg, Maria A1 - Hofmann, W.-K. A1 - Wernet, Peter A1 - Sieber, Reiner A1 - Müschen, Markus T1 - The BCR-ABL1 kinase bypasses selection for the expressio of a pre-B cell receptor in pre-B acute lymphoblastic leukemia cells Y1 - 2004 ER - TY - JOUR A1 - Frisch, D. A1 - Santer, B. T1 - Temperature-induced responses of a permanent-pond and a temporary-pond cyclopoid copepod : a link to habitat predictability? N2 - Temporary-pond species can be expected to use environmental cues to predict the onset of adverse conditions, while permanent-pond species may be insensitive to such cues. Temperature is such a potential cue in temporary waterbodies, as if fluctuates more widely with decreasing pond size than in deeper permanent ponds. We compared the temperature-induced response of a permanent-pond and a temporary-pond cyclopoid copepod focusing on juvenile development duration, diapause induction and survival during diapause. Nonlinear regression analysis suggested a stronger effect of temperature on the duration of juvenile development in the temporary-pond species. This species also showed a higher and temperature-dependent variation in development duration (highest coefficient of variation 26%) compared with the permanent species, for which variation was lower and similar at all temperatures (maximal coefficient of variation 6%). Temperature significantly influenced the induction of diapause in the temporary-pond species, where the percentage of individuals entering diapause increased from 0% at 5degreesC and 10degreesC to 63% at 15degreesC and 91% at 20degreesC. In the permanent-pond species, diapause induction was independent of temperature and was induced in 100% of experimental specimens. This suggests an obligatory diapause in the permanent-pond species, a type of dormancy that has not been described previously for cyclopoid copepods. Survival during diapause in both species was higher when the diapausing copepodid stage was reached at lower temperatures. At higher temperatures, the temporary-pond species survived longer than the permanent-pond species. These results suggest different temperature optima of the two species. The strategy displayed by the permanent-pond species might be selected for in more stable habitats and may preclude the colonization of temporary ponds. Higher flexibility in life-history traits and the use of temperature as an environmental cue in the temporary-pond species could be favoured in unpredictable habitats Y1 - 2004 SN - 1522-0613 ER - TY - JOUR A1 - Vaaje-Kolstad, G. A1 - Houston, Douglas R. A1 - Rao, F. V. A1 - Peter, Martin G. A1 - Synstad, Bjoenar A1 - van Aalten, Daan M. F. A1 - Eijsink, Vincent G. H. T1 - Structure of the D142N mutant of the family 18 chitinase ChiB from Serratia marcescens and its complex with allosamidin N2 - Catalysis by ChiB, a family 18 chitinase from Serratia marcescens, involves a conformational change of Asp142 which is part of a characteristic D140XD142XE144 sequence motif In the free enzyme Asp142 points towards Asp140, whereas it rotates towards the catalytic acid, Glu144, upon ligand binding. Mutation of Asp142 to Asn reduced k(cat) and affinity for allosamidin, a competitive inhibitor. The X-ray structure of the D142N mutant showed that Asn142 points towards Glu144 in the absence of a ligand. The active site also showed other structural adjustments (Tyr10, Ser93) that had previously been observed in the wild-type enzyme upon substrate binding. The X-ray structure of a complex of D142N with allosamidin, a pseudotrisaccharide competitive inhibitor, was essentially identical to that of the wild-type enzyme in complex with the same compound. Thus, the reduced allosamidin affinity in the mutant is not caused by structural changes but solely by the loss of electrostatic interactions with Asp142. The importance of electrostatics was further confirmed by the pH dependence of catalysis and allosamidin inhibition. The pH-dependent apparent affinities for allosamidin were not correlated with k(cat), indicating that it is probably better to view the inhibitor as a mimic of the oxazolinium ion reaction intermediate than as a transition state analogue. (C) 2003 Elsevier B.V. All rights reserved Y1 - 2004 SN - 1570-9639 ER -