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11.
12.
Isolation and characterization of two cDNA clones encoding ATP-sulfurylases from potato by complementation of a yeast mutant 总被引:3,自引:0,他引:3
Dirk Klonus Rainer Höfgen Lothar Willmitzer Jörg W. Riesmeier 《The Plant journal : for cell and molecular biology》1994,6(1):105-112
Sulfur plays an important role in plants, being used for the biosynthesis of amino acids, sulfolipids and secondary metabolites. After uptake sulfate is activated and subsequently reduced to sulfide or serves as donor for sulfurylation reactions. The first step in the activation of sulfate in all cases studied so far is catalyzed by the enzyme ATP-sulfurylase (E.C. 2.7.7.4.) which catalyzes the formation of adenosine-5′-phosphosulfate (APS). Two cDNA clones from potato encoding ATP-sulfurylases were identified following transformation of a Saccharomyces cerevisiae mutant deficient in ATP-sulfurylase activity with a cDNA library from potato source leaf poly(A)+ RNA cloned in a yeast expression vector. Several transformants were able to grow on a medium with sulfate as the only sulfur source, this ability being strictly linked to the presence of two classes of cDNAs. The clones StMet3-1 and StMet3-2 were further analyzed. DNA analysis revealed an open reading frame encoding a protein with a molecular mass of 48 kDa in the case of StMet3-1 and 52 kDa for StMet3-2. The deduced polypeptides are 88% identical at the amino acid level. The clone StMet3-2 has a 48 amino acid N-terminal extension which shows common features of a chloroplast transit peptide. Sequence comparison of the ATP-sulfurylase Met3 from Saccharomyces cerevisiae with the cDNA StMet3-1 (StMet3-2) reveals 31% (30%) identity at the amino acid level. Protein extracts from the yeast mutant transformed with the clone StMet3-1 displayed ATP-sulfurylase activity. RNA blot analysis demonstrated the expression of both genes in potato leaves, root and stem, but not in tubers. To the best of the authors' knowledge this is the first cloning and identification of genes involved in the reductive sulfate assimilation pathway from higher plants. 相似文献
13.
James B. Kramer Diane H. Boschelli David T. Connor Catherine R. Kostlan Paul J. Kuipers John A. Kennedy Clifford D. Wright Dirk A. Bornemeier Richard D. Dyer 《Bioorganic & medicinal chemistry letters》1993,3(12):2827-2830
The preparation of a series of 1,3,4-thiadiazoles and 1,3,4-oxadizoles linked by a thioether to 2,6-di-t-butylphenol and the inhibition of cyclooxygenase (CO) and 5-lipoxygenase (5-LO) by these compounds is dicussed. 相似文献
14.
Dirk T. Witte Frank J. Bruggeman Jan Piet Franke Swier Copinga Johanna M. Jansen Rokus A. De Zeeuw 《Chirality》1993,5(7):545-553
Direct enantiomeric separations of 17 chiral amidotetralins by means of high performance liquid chromatography were performed on stationary phases composed of tris(3,5-dimethylphenylcarbamate) derivatives of cellulose and amylose, coated on silica gel. The enantiomers of 15 out of 17 amidotetralins were resolved with a resolution of more than 1.5 by at least one of the chiral stationary phases. The stationary phases showed complementary results with regard to the separation of the amidotetralins, that is, pairs that did not separate on the cellulose-type column were well separated on the amylose-type column, and vice versa. There was no significant correlation between the chromatographic properties of the chiral stationary phases. © 1993 Wiley-Liss, Inc. 相似文献
15.
Molecular characterization of the proteinase-encoding gene, prb1, related to mycoparasitism by Trichoderma harzianum 总被引:11,自引:0,他引:11
Roberto A. Geremia Gustavo H. Goldman Dirk Jacobs W. Ardrtes Silvia B. Vila Marc Van Montagu Alfredo Herrera-Estrella 《Molecular microbiology》1993,8(3):603-613
The soil fungus Trichoderma harzianum is a mycoparasitic fungus known for its use as a biocontrol agent of phytopathogenic fungi. Among other factors, Trichoderma produces a series of antibiotics and fungal cell wall-degrading enzymes. These enzymes are believed to play an important role in mycoparasitism. Among the hydrolytic enzymes, we have identified a basic proteinase (Prb1) which is induced by either autoclaved mycelia, fungal cell wall preparation or chitin; however, the induction does not occur in the presence of glucose. The proteinase was purified and biochemically characterized as a serine proteinase of 31 kDa and pl 9.2. Based on the sequence of three internal peptides, synthetic oligonudeotide probes were designed. These probes allowed subsequent isolation of a cDNA and its corresponding genomic clone. The deduced amino acid sequence indicates that the proteinase is synthesized as a pre-proenzyme and allows its classification as a serine proteinase. Northen analysis shows that the induction of this enzyme is due to an increase in the corresponding mRNA level. 相似文献
16.
Effects of glucose, glucose plus branched-chain amino acids, or placebo on bike performance over 100km 总被引:1,自引:0,他引:1
Madsen Klavs; Maclean Dave A.; Kiens Bente; Christensen Dirk 《Journal of applied physiology》1996,81(6):2644-2650
Madsen, Klavs, Dave A. MacLean, Bente Kiens, and DirkChristensen. Effects of glucose, glucose plus branched-chain aminoacids, or placebo on bike performance over 100 km. J. Appl. Physiol. 81(6): 2644-2650, 1996.This studywas undertaken to determine the effects of ingesting either glucose(trial G) or glucose plusbranched-chain amino acids (BCAA; trialB), compared with placebo (trialP), during prolonged exercise. Nine well-trained cyclists with a maximal oxygen uptake of 63.1 ± 1.5 mlO2 · min1 · kg1performed three laboratory trials consisting of 100 km of cycling separated by 7 days between each trial. During these trials, the subjects were encouraged to complete the 100 km as fast as possible ontheir own bicycles connected to a magnetic brake. No differences inperformance times were observed between the three trials (160.1 ± 4.1, 157.2 ± 4.5, and 159.8 ± 3.7 min, respectively). Intrial B, plasma BCAA levels increased from339 ± 28 µM at rest to 1,026 ± 62 µM after exercise(P < 0.01). Plasma ammoniaconcentrations increased during the entire exercise period for allthree trials and were significantly higher intrial B compared withtrials G andP (P < 0.05). The respiratory exchange ratio was similar in the threetrials during the first 90 min of exercise; thereafter, it tended todrop more in trial P than intrials G andB. These data suggest that neitherglucose nor glucose plus BCAA ingestion during 100 km of cyclingenhance performance in well-trained cyclists. 相似文献
17.
Plantains (Musa spp., AAB group) are an important food crop and an integral component of the farming systems in the lowland humid forest zone of West and Central Africa. A group of 24 plantain cultivars, representing the major variability in West Africa, was evaluated for nine quantitative characters. The association between growth and yield parameters in this African plantain germplasm was examined to determine if the pattern of quantitative variation in inflorescence and vegetative traits agreed with taxonomic groupings based on inflorescence type and plant size. Phenotypic correlations between these traits were calculated. Giant cultivars were taller, their pseudostem thicker, and they flowered much later than medium-sized cultivars. Giant cultivars produced more foliage, resulting in heavier bunches with more hands and fruits. Groupings that resulted following principal component analysis (PCA) supported conventional taxonomic groupings of plantains. PC A was based mainly on time to flowering, pseudostem height, and number of fruits. The last two traits, in combination with the number of hermaphrodite flowers and the persistence of the male bud, sufficed to group plantain cultivars. 相似文献
18.
Daniel Duprez Dirk Voet Marc De Buyzere Benny Drieghe Bruno Vyncke Sigrid Mareels Marcel Afschrift Denis L. Clement 《European journal of applied physiology and occupational physiology》1995,71(5):459-463
The splanchnic circulation can make a major contribution to blood flow changes. However, the role of the splanchnic circulation in the reflex adjustments to the blood pressure increase during isometric exercise is not well documented. The central command and the muscle chemoreflex are the two major mechanisms involved in the blood pressure response to isometric exercise. This study aimed to examine the behaviour of the superior mesenteric artery during isometric handgrip (IHG) at 30% maximal voluntary contraction (MVC). The pulsatility index (PI) of the blood velocity waveform of the superior mesenteric artery was taken as the study parameter. A total of ten healthy subjects [mean age, 21.1 (SEM 0.3) years] performed an IHG at 30% MVC for 90 s. At 5 s prior to the end of the exercise, muscle circulation was arrested for 90 s to study the effect of the muscle chemoreflex (post exercise arterial occlusion, PEAO). The IHG at 30% MVC caused a decrease in superior mesenteric artery PI, from 4.84 (SEM 1.57) at control level to 3.90 (SEM 1.07) (P = 0.015). The PI further decreased to 3.17 (SEM 0.70) (P = 0.01) during PEAO. Our results indicated that ergoreceptors may be involved in the superior mesenteric artery vasodilatation during isometric exercise. 相似文献
19.
20.
Mycorrhizal and nonmycorrhizal roots of Allium schoenoprasum were tested for activities of α-mannosidase, β-glucosidase and arabinosidase. Mannosidase activity was higher by a factor
of two in mycorrhizal than in nonmycorrhizal root extracts. The apparent molecular weight of the enzyme was 152 kDa and its
KM was 1.25 mM in colonized roots and 1.85 mM in uncolonized roots. α-Mannosidase activity was further characterized by an acid
pH optimum and Zn2+ dependency. No significant differences could be found between mycorrhizal and nonmycorrhizal roots for β-glucosidase and
arabinosidase activities.
Accepted: 28 August 1995 相似文献