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排序方式: 共有225条查询结果,搜索用时 15 毫秒
1.
2.
Solubilization and Purification of NAD(P)H Dehydrogenase of Cucurbita Microsomes 总被引:4,自引:4,他引:0 下载免费PDF全文
An NAD(P)H dehydrogenase stimulated by quinone (P Pupillo, V Valenti, L de Luca, R Hertel 1986 Plant Physiol 80: 384-389) was solubilized from washed microsomes of zucchini squash hypocotyls (Cucurbita pepo L.) by use of 1% Triton X-100. The solubilized enzyme remained in solution in aqueous buffer and could be purified by a combination of Sepharose 6B chromatography and Blue Ultrogel chromatography. Of the three peaks of activity eluted from the latter column with a salt gradient, peak 3 had 50% or more of the activity and was almost pure enzyme. The preparation examined in SDS-gel electrophoresis consisted of two types of subunits, a (molecular weight 39,500) and b (37,000) in equal amounts. Peak 2 was less pure but had a similar polypeptide pattern. The active protein is proposed to be a heterotetramer (a2b2) having a molecular weight of about 150,000, as found by gel exclusion chromatography. The purified enzyme can reduce several quinones, DCPIP, cytochrome c, and with best efficiency ferricyanide, and is therefore a diaphorase. The kinetics for the substrates are negatively cooperative with Hill coefficients nH = 0.55 ± 0.05 for NADPH and 0.22 ± 0.04 for duroquinone. A weak inhibition by p-hydroxymercuric benzoate and mersalyl (stronger with microsomal preparations) suggests the presence of essential sulfhydryl group(s). The possibility is discussed that the dehydrogenase is an NAD(P)H-P450 reductase or similar flavoprotein, and that it is responsible for the NADPH-cytochrome c reductase activity of plant microsomes. 相似文献
3.
G Bartolini M Orlandi M Chiricolo L Minghetti F Guerrini M Fidan C Franceschi V Tomasi 《Biochimica et biophysica acta》1986,876(3):486-493
It has previously been shown that platelet-free human monocytes, when properly incubated in the presence of animal and human sera, became capable of producing large amounts of thromboxane A2 and prostaglandin E2. The characteristics of these processes are reported here. Prostaglandin biosynthesis was time and cell concentration dependent; 24 h of incubation at 37 degrees C and 0.5 X 10(6) cells per ml medium were found to give the most reproducible results. Human monocytes produced thromboxane A2 and prostaglandin E2 in a typical ratio which ranged from 2.0 to 5.0 (28 experiments). Animal and human sera were similarly effective, while serum obtained from platelet-free blood was much less active. The activity of all sera tested was stable to heating (100 degrees C for 2-10 min) and extreme pH values (pH 2 and 11). It was unstable when the serum was heated at pH 11 and after 2-mercaptoethanol treatment. These observations prompted us to check the effect of polypeptide growth factors having properties similar to those reported above, such as platelet-derived growth factor, fibroblast growth factor, epidermal growth factor as well as insulin and transferrin. None of these, alone or in various combinations, was capable of eliciting a stimulation comparable with that of serum. Stimulation due to sera was, as expected, dose dependently inhibited by acetylsalicylic acid and more efficiently by indomethacin; unexpectedly it was also inhibited by protein synthesis inhibitors such as actinomycin D and cycloheximide in conditions under which no toxic effect of the drugs was evident. On the basis of these results we conclude that: (a) polypeptide growth factor(s) with a molecular weight at least 30 000 (as judged by Amicon ultrafiltration) is involved in the regulation of prostaglandin biosynthesis); (b) such a factor(s) acts by inducing rather than by activating the cyclooxygenase system. 相似文献
4.
Guerrini Franca; Lombini Alessandra; Bizarri Mariachiara; Pupillo Paolo 《Journal of experimental botany》1994,45(9):1227-1233
Cell suspension cultures of Beta vulgaris L., treated with calciumchelators or untreated, were used to characterize pyndine nucleotide-dependentdiaphorases of microsomes. The microsomal activity of NADH-dependentduroquinone reductase from cultures treated with 10 mM Na2EGTAfor 24 h increased by a factor of 1.8 with respect to controlmicrosomes, and was mainly associated with particles of d=1.17gml1. NADPH-duroquinone reductase and NADH-ferricyanidereductase activities showed smaller increases. Bacterial protein-lipopolysaccharidecomplexes (prLPS) also promoted the increase of microsomal diaphorases;CaEGTA was Ineffective. EGTA effects on enzymes of supernatantand mitochondria were negligible, although Na2EGTA treatmentinduced cell aggregation and strong acidification of the medium. When microsomes from control cultures were solubilized with1% LPC and fractionated in high-efficiency gel permeation columns(FPLC) the diaphorase activities were found associated to threemajor proteins: (i) NADH-specific quinone reductase (NADH-QR)of 340 kDa; (ii) pyndine nucleotide-nonspecific quinone reductase(NAD(P)H-QR) of 85 kDa also having ferricyanide reductase activity;(iii) NADH-specific ferricyanide reductase (NADH-FCR) of 38kDa. The microsomes from EGTA-treated cells also showed a highlyactive NADH-QR having a larger molecular mass (440 kDa) thanin control cells. NAD(P)H-QR also showed increased activity.We conclude that external Ca2+ chelation induces changes indehydrogenase components in microsomes. Furthermore, prLPS probablyexert part of their effect on plants through Ca2+ chelation. Key words: Beta vulgaris, cell cultures, calcium chelators, diaphorase, NAD(P)H-dehydrogenase, lipopolysaccharide, EGTA, quinone reductase 相似文献
5.
S. Salvadori R. Guerrini V. Forlani S. D. Bryant M. Attila L. H. Lazarus 《Amino acids》1994,7(3):291-304
Summary Analysis of deltorphin A position 4 analogues included: backbone constrained N
MeHis, spinacine (Spi), N
MePhe and the tetrahydroisoquinoline-3-carboxylic acid (Tic); spatially confined side-chain (Phg); and imidazole alkylation ofl- andd-His4 enantiomers. High selectivity was lost with the following replacements: N
MeHis4, N
MePhe4 and Phg4 reduced binding and the constrained residues also increasedµ binding; ring closure between the side-chain and amino group to yield Spi4 or Tic4 increasedµ affinity. Imidazole methylation of His4 marginally affected opioid binding and doubled selectivity; alkylatedd-His4-derivatives generally maintained selectivity in spite of decreased affinities. Thus, His4 imidazole preserves selectivity by facilitating high binding and by repulsion at theµ receptor. Several low energy conformers of deltorphin A indicated that the His4 imidazole preferred a spatial orientation parallel to the phenolic side-chain of Tyr1 suggestive that this conformation might contribute to high affinity and selectivity. 相似文献
6.
A Casino M Cipollaro A M Guerrini G Mastrocinque A Spena V Scarlato 《Nucleic acids research》1981,9(6):1499-1518
A Fortran computer algorithm has been used to analyze the nucleotide sequence of several structural genes. The analysis performed on both coding and complementary DNA strands shows that whereas open reading frames shorter than 100 codons are randomly distributed on both DNA strands, open reading frames longer than 100 codons ("virtual genes") are significantly more frequent on the complementary DNA strand than on the coding one. These "virtual genes" were further investigated by looking at intron sequences, splicing points, signal sequences and by analyzing gene mutations. On the basis of this analysis coding and complementary DNA strands of several eukaryotic structural genes cannot be distinguished. In particular we suggest that the complementary DNA strand of the human epsilon-globin gene might indeed code for a protein. 相似文献
7.
Summary The growth of two strains of the petitenegative yeast Kluyveromyces lactis is inhibited by Tetracycline in different ways under the same culture conditions. Tetracycline resistant mutants of one strain have been isolated which can tolerate doses as high as 3000 g/ml of antibiotic.The segregation pattern of this character obtained by random spore analysis of the ascospores derived from the cross of the two strains strongly suggests that the resistance to tetracycline is under mitochondrial control. 相似文献
8.
Corrections to: Apparent recruitment failure for the vast majority of coral species at Eilat,Red Sea
Guerrini Gabriele Yerushalmy Maayan Shefy Dor Shashar Nadav Rinkevich Baruch 《Coral reefs (Online)》2021,40(4):1393-1395
Coral Reefs - A correction to this paper has been published: https://doi.org/10.1007/s00338-021-02121-x 相似文献
9.
Phytotoxic Effects and Phytochemical Fingerprinting of Hydrodistilled Oil,Enriched Fractions,and Isolated Compounds Obtained from Cryptocarya massoy (Oken) Kosterm. Bark 下载免费PDF全文
Enrico Rolli Matteo Marieschi Silvia Maietti Alessandra Guerrini Alessandro Grandini Gianni Sacchetti Renato Bruni 《化学与生物多样性》2016,13(1):66-76
The hydrodistilled oil of Cryptocarya massoy bark was characterized by GC‐FID and GC/MS analyses, allowing the identification of unusual C10 massoia lactone ( 3 , 56.2%), C12 massoia lactone ( 4 , 16.5%), benzyl benzoate ( 1 , 12.7%), C8 massoia lactone (3.4%), δ‐decalactone ( 5 , 1.5%), and benzyl salicylate ( 2 , 1.8%) as main constituents. The phytotoxic activities of the oil, three enriched fractions (lactone‐rich, ester‐rich, and sesquiterpene‐rich), and four constituents (compounds 1, 2, 5 , and δ‐dodecalactone ( 6 )) against Lycopersicon esculentum and Cucumis sativus seeds and seedlings were screened. At a concentration of 1000 μl/l, the essential oil and the massoia lactone‐rich fraction caused a complete inhibition of the germination of both seeds, and, when applied on tomato plantlets, they induced an 85 and 100% dieback, respectively. These performances exceeded those of the well‐known phytotoxic essential oils of Syzygium aromaticum and Cymbopogon citratus, already used in commercial products for the weed and pest management. The same substances were also evaluated against four phytopathogenic bacteria and ten phytopathogenic fungi, providing EC50 values against the most susceptible strains in the 100–500 μl/l range for the essential oil and in the 10–50 μl/l range for compound 6 and the lactone‐rich fraction. The phytotoxic behavior was related mainly to massoia lactones and benzyl esters, while a greater amount of 6 may infer a good activity against some phytopathogenic fungi. Further investigations of these secondary metabolites are warranted, to evaluate their use as natural herbicides. 相似文献
10.
Gabriella Guerrini Giovanna Ciciani Samuele Ciattini Letizia Crocetti Simona Daniele Claudia Martini 《Journal of enzyme inhibition and medicinal chemistry》2016,31(2):195-204
To investigate the binding affinity of GABAA receptor subtype, new pyrazolo [1,5-a]quinazolines were designed, synthesized, and in vitro evaluated. These compounds, 5-deaza analogues of pyrazolo[5,1-c][1,2,4]benzotriazine derivatives which were already studied in our research group, permit us to evaluate the relevance of the nitrogen or the oxygen atom at 5-position of the tricyclic scaffold. Molecular dynamic study was done on a set of the new and known ligands to rationalize and to explain the lack of affinity on the 4- or 5-substituted new derivative. In fact, from biological results, it can be found that the only 5-unsubstituted new derivative, compound 15, has receptor recognition (Ki?=?834.7?nM). 相似文献