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1.
As part of a program to explore the structural requirement of N-glycans in the carbohydrate-mediated biological interactions, N-linked pentasaccharide core structure was stereochemically modified in terms of glycosidic linkage. Three isomers, -D-Man-(13)-[-D-Man-(16)]--D-Man-(14)--D-GlcNAc-(14)--D-GlcNAc-L-Asn, -D-Man-(13)-[-D-Man-(16)]--D-Man-(14)--D-GlcNAc-(14)--D-GlcNAc-L-Asn, and -D-Man-(13)-[-D-man-(16)]--D-Man-(14)--D-GlcNAc-(14)--D-GlcNAc-L-Asn, were synthesized. Synthesis of the pentasaccharide with natural linkage is also described.  相似文献   

2.
1. Opioid (, , ) and cannabinoid (CB1, CB2) receptors are coupled mainly toGi/Go GTP-binding proteins. The goal of the present study was to determine whether different subtypes of opioid and cannabinoid receptors, when coexpressed in the same cell, share a common reservoir, or utilize different pools, of G proteins.2. The stimulation of [35S]GTPS binding by selective opioid and cannabinoid agonists was tested in transiently transfected COS-7 cells, as well as in neuroblastoma cell lines. In COS-7 cells, cotransfection of - and -opioid receptors led to stimulation of [35S]GTPS binding by either -selective (DAMGO) or -selective (DPDPE) agonists. The combined effect of the two agonists was similar to the effect of either DAMGO or DPDPE alone, suggesting the activation of a common G-protein reservoir by the two receptor subtypes.3. The same phenomenon was observed when COS-7 cells were cotransfected with CB1 cannabinoid receptors and either - or -opioid receptors.4. On the other hand, in N18TG2 neuroblastoma cells, which endogenously coexpress CB1 and -opioid receptors, as well as in SK-N-SH neuroblastoma cells, which coexpress - and -opioid receptors, the combined effects of the various agonists (the selective cannabinoid DALN and the selective opioids DPDPE and DAMGO) were additive, implying the activation of different pools of G proteins by each receptorsubtype.5. These results suggest a fundamental difference between native and artificially transfected cells regarding the compartmentalization of receptors and GTP-binding proteins.  相似文献   

3.
C. Duffus  R. Rosie 《Planta》1973,109(2):153-160
Summary The enzymes -amylase (-1, 4-glucan 4-glucanohydrolase, 3.2.1.1), -amylase (-1,4-glucan maltohydrolase, 3.2.1.2) and phosphorylase (-1,4-glucan: orthophosphate glucosyltransferase, 2.4.1.1) were assayed in whole grains of barley throughout the maturation period. -amylase and phosphorylase had peaks of activity between 25 and 30 days after anthesis. On the other hand the activity of -amylase in both the available and latent forms reached a maximum value at 35 days after anthesis which did not decrease thereafter. -amylase activity was also assayed throughout development in the endosperm, aleurone, testa pericarp and embryo. Latent -amylase reached a constant maximum value in endosperm at 35 days but available -amylase reached a peak of activity at 25 days and then declined to zero at 45 days. Only latent -amylase was associated with the aleurone layer and activity rose to a maximum value at 35 days. The testa pericarp had mainly latent -amylase whose activity fell from an early maximum at 21 days to zero at 35 days. No hydrolytic activity was associated with the embryo. The phosphorylase activity was low and mainly associated with the endosperm fraction.  相似文献   

4.
Summary The fluorescent intercalation complex of ethidium bromide (ETB) with DNA was used as a probe to compare the effects of various radicals with respect to impairment of the DNA base-pair region.OH radicals inhibit up to 0.7 dye intercalations perOH at low salt concentration, and for various oxidizing species the effect decreases in the orderOH > Br 2 > N 3 > $$ " align="middle" border="0"> (SCN) 2 . DNA impairment by theOH product of Met-Gly is comparable to that of N 3 , but no effect was found due to the interaction between DNA and Lys-Tyr-Lys phenoxyl radicals. The reducing speciese aq , H, O 2 , and CO 2 hardly affect the DNA-ETB intercalation.  相似文献   

5.
Summary Thirteen maize (Zea mays L.) populations including five adapted, five adapted x exotic, two composites of adapted and exotic, and one exotic selected for adaptability were crossed in a diallel mating system. The parents and 78 crosses and nine check hybrids were evaluated for grain yield and plant height in five environments. The Gardner-Eberhart model Analysis II indicated that additive and nonadditive gene effects accounted for 60 and 40% of the total variation among populations, respectively, for grain yield and 86% and 14% of the total variation, respectively, for plant height. Components of heterosis were significant in the combined analysis for both traits. Adapted Corn Belt populations tended to have higher performance in crosses and greater values of variety heterosis than 50% adapted populations. Nebraska Elite Composite, Corn Belt x Mexican, and Corn Belt x Brazilian showed high mean yields in crosses, however, they were not among those with high estimates of variety heterosis. One exotic population (Tuxpeno x Antigua Grupo 2) and three adapted populations [307 Composite, NB(S1)C-3, and NK(S1)C-3] might be combined together to form a high-yielding population. It may be possible to synthesize two useful populations for reciprocal recurrent selection by grouping Tuxpeno x Antiqua Grupo 2, NB(S1)C-3, and NS(FS)LFW-8 into one population and NK(S1)C-3, KrugxTabloncillo, and 307 Composite in the other one.Contribution from the Department of Agronomy, University of Nebraska, Lincoln, NE 68583. Published as Paper No. 8011. Journal Series, Nebr. Agric. Exp. Station. Research was conducted under Project 12-049  相似文献   

6.
Summary An actinomycete (MA 6474, ATCC 53828) isolated from a soil sample (Mutare, Zimbabwe) was found to biotransform the sodium salt of Simvastatin (MK-733) to 6--hydroxymethyl MK-733, 6--hybroxymethyl MK-733, and 6-ring-hydroxy MK-733. The bioconversion efficiency to the desired compound, 6--hydroxymethyl MK-733, was enhanced by optimizing the physico-chemical parameters of the process. In shake flask cultures, addition of magnesium (0.125 mg/l Mg SO4·7H2O) to the medium resulted in a five-fold increase in the rate of bioconversion to the diastereomer. The ratio of bioconversion products (6--hydroxymethyl, 6--hydroxymethyl, and 6-ring-hydroxy MK-733) was regulated by pH. Process improvements and scale up in 23-1 fermentors, which consisted of a controlled addition of substrate (MK-733), resulted in a 2-fold increase in alpha diastereomer Production (42 vs. 79 U/ml) and a 23-fold rate increase in the formation of -diastereomer. A high diastereomeric ratio (: =91) facilitated downstream processing.  相似文献   

7.
The time constant of the process producing the delay in Na inactivation development as determined by the two pulse method (delay) was extracted and compared to that of the slowest Na activation process 3 for the I Na during the conditioning pulse of that same determination. delay and two pulse inactivation c values were computer generated using a nonlinear least squares algorithm. h and single pulse inactivation h values were independently generated for each determination also with the aid of the computer using the same non-linear least squares algorithm. In one determination at 2 mV, c was 4.68 and delay 0.494 ms while h was 4.70 and 3 0.491 ms for a c/h of 0.996 and a delay/3 of 1.006. Mean delay/3 from five determinations in four axons, both Cs and K perfused, and spanning a potential range of-27 to 2mV was 1.068. The precursor process to inactivation is channel opening. Some fraction of channels presumably inactivate via another route where prior channel opening is not required.  相似文献   

8.
The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update   总被引:14,自引:0,他引:14  
Mitochondrial dysfunction has been shown to participate in the induction of apoptosis and has even been suggested to be central to the apoptotic pathway. Indeed, opening of the mitochondrial permeability transition pore has been demonstrated to induce depolarization of the transmembrane potential (m), release of apoptogenic factors and loss of oxidative phosphorylation. In some apoptotic systems, loss of m may be an early event in the apoptotic process. However, there are emerging data suggesting that, depending on the model of apoptosis, the loss of m may not be an early requirement for apoptosis, but on the contrary may be a consequence of the apoptotic-signaling pathway. Furthermore, to add to these conflicting data, loss of m has been demonstrated to not be required for cytochrome c release, whereas release of apoptosis inducing factor AIF is dependent upon disruption of m early in the apoptotic pathway. Together, the existing literature suggests that depending on the cell system under investigation and the apoptotic stimuli used, dissipation of m may or may not be an early event in the apoptotic pathway. Discrepancies in this area of apoptosis research may be attributed to the fluorochromes used to detect m. Differential degrees of sensitivity of these fluorochromes exist, and there are also important factors that contribute to their ability to accurately discriminate changes in m.  相似文献   

9.
Narcolepsy has a 98% association with the DR2-Dw2/DQw1 haplotype. To establish if a disease-specific allele is present in narcolepsy, a cDNA library was made from a B-cell line from a DR2,4/DQw1,3 narcoleptic. Clones encoding the two expressed DR2 chains, along with DQw1 and chains, were isolated and completely sequenced. The coding regions of these four genes were similar to published nucleotide and protein sequences from corresponding healthy controls, with some minor exceptions. The 3 untranslated region of one of the DR2 genes in the narcoleptic was extended by 42 bp. Complete sequences were not available for DQw1.2 or from healthy individuals, but first domain nucleotide sequences showed only a single nonproductive difference in DQ. Partial protein sequences of both DQ and from published data were identical. Although the effects of minor differences cannot be ruled out completely, it is concluded that there are probably no narcolepsy-specific DR or DQ / sequences, and that the alleles found in narcolepsy are representative of those found in the healthy population.  相似文献   

10.
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.  相似文献   

11.
Summary 2-Amino-2-deoxyuridine reacts efficiently with nucleoside 5-phosphorimidazolides in aqueous solution. The dinucleoside monophosphate analogues were obtained in yields exceeding 80% under conditions in which little reaction occurs with the natural nucleosides.In a similar way, the 5-phosphorimidazolide of 2-amino-2-deoxyuridine undergoes self-condensation in aqueous solution to give a complex mixture of oligomers.The phosphoramidate bond in the dinucleoside monophosphate analogues is stable for several days at room temperature and pH 7. The mechanisms of their hydrolysis under acidic and alkaline conditions are described.Abbreviations A adenosine - C cytidine - G guanosine - U uridine - T thymidine - UN 3 2-azido-2-deoxyuridine - UNH 2 2-amino-2-deoxyuridine - ImpA adenosine 5-phosphorimidazolide - ImpU uridine 5-phosphorimidazolide - ImpUN 3 2-azido-2-deoxyuridine 5-phosphorimidazolide - ImpUNH 2 2-amino-2-deoxyuridine 5-phosphorimidazolide - pA adenosine 5-phosphate - pU uridine 5-phosphate - pUN 3 2-azido-2-deoxyuridine 5-phosphate - pUNH 2 2-amino-2-deoxyuridine 5-phosphate - UpA uridylyl-[35]-adenosine - UpU uridylyl-[35]-uridine - UNpA adenylyl-[52]-2-amino-2-deoxy-uridine - UNpU uridylyl-[52]-2-amino-2-deoxyuridine (pUN)n n=2,3,4 [25]-linked oligomers of pUNH 2 poly(A) polyadenylic acid - Im imidazole - MeIm l-methylimidazole  相似文献   

12.
Summary Ascorbate has been related to the differentiation of several mesenchymal cells including haematopoietic cells. We have previously demonstrated that ascorbate enhances the activity of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on monocytic differentiation of HL-60 cells. Here, we show that ascorbate-mediated modification of cellular redox state and AP-1 (activating protein-1) DNA binding during early phases are related to the enhancing effect of ascorbate on differentiation. Ascorbate, but not its fully oxidized form, dehydroascorbate, or an ascorbate analogue with a low rate of oxidation, ascorbate-2-phosphate, enhanced the differentiation induced by 1,25(OH)2D3, modified cytosolic reactive oxygen species levels and mitochondrial redox potential (m), and modulated AP-1 DNA binding in HL-60 cells. Ascorbate itself increased AP-1 binding to DNA in noninduced cells, whereas it inhibited AP-1 binding in 1,25(OH)2D3-induced cells. However, ascorbate increased the mRNA levels ofc-jun, junB. andc-fos in 1,25(OH)2D3-induced cells. Taken together, these results suggest that the enhancing effect of ascorbate on HL-60 differentiation induced by 1,25(OH)2D3 is related to its effect on the cellular redox state and the modulation of AP-1 activity.Abbreviations 1,25-(OH)2D3 1,25-dihydroxyvitamin D3 - m mitochondrial transmembrane potential - AP-1 activating protein-1 - Asc-2-P ascorbate-2-phosphate - DHA dehydroascorbate - DiOC6(3) 3,3-dihexyloxacarboxyanine iodide - EMSA electromobility shift assay - NBT nitroblue tetrazolium - ROS reactive oxygen species  相似文献   

13.
Based on different marker information content mapping of QTLs for Fusarium head blight resistance in wheat was compared with regard to number and consistency of detected QTLs as well as QTL positions and effects. Therefore, two linkage maps, obtained by dominant and codominant genotyping of hemizygous markers, were constructed with 211 AFLPs, 37 SSRs and the barley RGA marker XaACT/CAA. The codominant marker set comprised 59% codominant markers, whereas the dominant map consisted of only 13%. A segregating wheat population of 94 F4-RILs was used for QTL analysis. Fusarium head blight resistance was estimated in field trials in six environments. Conventional dominant marker scoring found seven QTLs. The phenotypic variations explained by QTLs detected in single environment analyses ranged from 11.1 to 44.6%. QTL analysis performed with the codominant marker set confirmed not only all QTL positions as revealed by dominant QTL analysis', but also 12 additional QTLs were found. QTLs in single environments explained 36.3 up to 55.7% of the phenotypic variation. In the QTL analysis across all environments, none of the QTLs could be confirmed using dominant marker scoring. However, by codominant QTL analysis' environment-specific QTLs were retrieved. STS marker XaACT/CAA was found to be significantly associated with FHB resistance only by codominant scoring. Support intervals of QTLs commonly found in both marker sets averaged to 10.3 cM in the dominant QTL analysis', whereas the length was shortened to 8.9 cM by codominant genotyping. The advantages of extracting codominant information from dominant markers are discussed.  相似文献   

14.
Summary The zymogram phenotypes that Chinese Spring-Imperial, Holdfast-King II and Kharkov-Dakold wheat-rye addition lines presented for esterase isozymes were determined using polyacrylamide gel ectrophoresis. The analyses were carried out with different parts of the dry kernel, namely embryo plus scutellum and endosperm, leaves and roots. In all cases, embryo plus scutellum, endosperm and leaf presented different patterns of esterases. The patterns of leaves and roots were the same. Results indicate that rye esterases exist as monomers and dimers. Dimeric esterases are controlled by one locus located on the 3R chromosomes of Imperial, King II and Dakold rye cultivars. Five loci involved in the production of monomeric esterases have been located on the 6R chromosomes of these cultivars, specifically on the long arm of the King II 6R chromosome. On the basis of these results, considerations concerning chromosome homoeology and homology are made.  相似文献   

15.
Crude extracts of Rhodospirillum rubrum catalyzed the formation of acid-volatile radioactivity from (35S) sulfate, (35S) adenosine-5-phosphosulfate, and (35S) 3-phosphoadenosine-5-phosphosulfate. An enzyme fraction similar to APS-sulfotransferases from plant sources was purified 228-fold from Rhodospirillum rubrum. It is suggested here that this enzyme is specific for adenosine-5-phosphosulfate, because the purified enzyme fraction metabolized adenosine-5-phosphosulfate, however, only at a rate of 1/10 of that with adenosine-5-phosphosulfate. Further, the reaction with 3-phosphoadenosine-5-phosphosulfate was inhibited with 3-phosphoadenosine-5-phosphate whereas this nucleotide had no effect on the reaction with adenosine-5-phosphosulfate. For this activity with adenosine-5-phosphosulfate the name APS-sulfotransferase is suggested. This APS-sulfotransferase needs thiols for activity; good rates were obtained with either dithioerythritol or reduced glutathione; other thiols like cysteine, 2-3-dimercaptopropanol or mercaptoethanol are less effective. The electron donor methylviologen did not catalyze this reaction. The pH-optimum was about 9.0; the apparent K m for adenosine-5-phosphosulfate was determined to be 0.05 mM with this so far purified enzyme fraction. Enzyme activity was increased with K2SO4 and Na2SO4 and was inhibited by 5-AMP. These properties are similar to assimilatory APS-sulfotransferases from spinach and Chlorella.Abbreviations APS adenosine-5-phosphosulfate - PAPS 3-phosphoadenosine-5-phosphosulfate - 5-AMP adenosine-5-monophosphate - 3-AMP adenosine-3-monophosphate - 3-5-ADP 3-phosphoadenosine-5-phosphate (PAP) - DTE dithiorythritol - GSH reduced glutathione - BAL 2-3-dimercaptopropanol  相似文献   

16.
Dry lettuce (Lactuca sativa L.) seeds (achenes) contain -galactosidase (EC 3.2.122) at a level which is maintained in the imbibed dormant state in darkness. Both red light (R) and gibberellic acid promote an increase in enzyme activity several hours prior to the completion of germination. Germination and enzyme activity are not essentially linked, however, for the latter can increase while the former is inhibited. -Galactosidase activity increases within the cotyledons and the endosperm following R stimulation, but the axis is essential to perceive the stimulus and to promote and maintain the increase in enzyme activity. A diffusible factor (or factors) is produced by and-or released from irradiated axes, and it migrates to the cotyledons (and possibly endosperm) to promote the increase in -galactosidase activity. Gibberellic acid, particularly in the presence of benzyladenine, can replace the requirement for irradiated axes.Abbreviations GA3 gibberellic acid - R red light  相似文献   

17.
The relative roles of the two structural aspects of nonenzymic glycation sites of hemoglobin A, namely the ease with which the amino groups could form the aldimine adducts and the propensity of the microenvironments of the respective aldimines to facilitate the Amadori rearrangement, in dictating the site selectivity of nonenzymic glycation with aldotriose has been investigated. The chemical reactivity of the amino groups of hemoglobin A forin vitro reductive glycation with aldotriose is distinct from that in the nonreductive mode. The reactivity of amino groups of hemoglobin A toward reductive glycation (i.e., propensity for aldimine formation) decreases in the order Val-1(), Val-1(), Lys-66(), Lys-61(), and Lys-16(). The overall reactivity of hemoglobin A toward nonreductive glycation decreased in the order Lys-16(), Val-1(), Lys-66(), Lys-82(), Lys-61(), and Val-1(). Since the aldimine is the common intermediate for both the reductive and nonreductive modification, the differential selectivity of protein for the two modes of glycation is clearly a reflection of the propensity of the microenvironments of nonenzymic glycation sites to facilitate the isomerization reaction (i.e., Amadori rearrangement). A semiquantitative estimate of this propensity of the microenvironment of the nonenzymic glycation sites has been obtained by comparing the nonreductive (nonenzymic) and reductive modification at individual glycation sites. The microenvironment of Lys-16() is very efficient in facilitating the rearrangement and the relative efficiency decreases in the order Lys-16(), Lys-82(), Lys-66(), Lys-61(), Val-1(), and Val-1(). The propensity of the microenvironment of Lys-16() to facilitate the Amadori rearrangement of the aldimine is about three orders of magnitude higher than that of Val-1() and is about 50 times higher than that of Val-1(). The extent of nonenzymic glycation at the individual sites is modulated by various factors, such as thepH, concentration of aldotriose, and the concentration of the protein. The nucleophiles—such as tris, glycine ethyl ester, and amino guanidine—inhibit the glycation by trapping the aldotriose. The nonenzymic glycation inhibitory power of nucleophile is directly related to its propensity to form aldimine. Thus, the extent of inhibition of nonenzymic glycation at a given site by a nucleophile directly reflects the relative role ofpK a of the site in dictating the glycation at that site. The nonenzymic glycation of an amino group of a protein is an additive/synergestic consequence of the propensity of the site to form aldimine adducts on one hand, and the propensity of its microenvironment to facilitate the isomerization of the aldimines to ketoamines on the other. The isomerization potential of microenvironment plays the dominant role in dictating the site specificity of the nonenzymic glycation of proteins.  相似文献   

18.
F1-ATPases are large multimeric proteins that can be isolated from the membrane bound system that catalyzes the phosphorylation of ADP by inorganic phosphate in bacteria, plants, and mitochondria. They can be visualized in electron micrographs of the inner mitochondrial membranes where they appear as large protruding spheres 90 Å in diameter. The purified F1-ATPases have a molecular weight of 320,000 to 400,000 daltons and are composed of five non-identical subunits (, , , and ). The stoichiometry of these subunits in the complex is still unknown but compositions of the type 33 and 22222 were found to be consistent with some of the available experimental data. This review discusses the recent data and the experimental approaches utilized for the structural characterization of F1-ATPases.  相似文献   

19.
Summary Dominant alleles of the Kr1 and Kr2 genes reduce the crossability of hexaploid wheat with many alien species, including rye and Hordeum bulbosum, with Kr1 having the greater effect. However, a cytological study of wheat ovaries fixed 48 h after pollination showed that the wheat genotypes Highbury (kr1, Kr2) and Chinese Spring (Hope 5B) (kr1, kr2) were crossable with Seneca 60 maize, fertilization occurring in 14.4 and 30.7% of embryo sacs respectively. The latter figure was similar to the 29.7% fertilization found in Chinese Spring (kr1, kr2). Most embryo sacs in which fertilization occurred contained an embryo but lacked an endosperm and where an endosperm was formed it was usually highly aberrant. All three wheat x maize combinations were karyotypically unstable and rapidly eliminated maize chromosomes to produce haploid wheat embryos.  相似文献   

20.
Peptide acceptor properties of phenylalanine and glycine esters of 9-(2,3-dihydroxypropyl-1)-adenine and 1-(2,3-dihydroxypropyl-1)-4-thiouracyl were investigated. All these esters appeared to be powerful inhibitors of polyphenylalanine synthesis in E. coli MRE-600 ribosomes charged with poly U. Like puromycin, esters of adenine derivatives accepted the AcPhe residue from Ac-[14C] Phe-tRNA in a ribosomal system charged with poly U. However, peptidyl esters of 9-(2,3-dihydroxypropyl-1)-adenine remained bound with ribosomes. The structure of the peptide esters synthesized was ascertained after dissociation of ribosomes into subparticles by direct comparison with the synthetic specimens.Abbreviations AcPhe acetyl-l-phenylalanine - HP-Ade 9-(2,3-dihydroxypropyl-1)-adenine - Phe-HP-Ade and Gly-HP-Ade l-phenylalanine and glycine esters of HP-Ade - Phe-HP-TUra l-phenylalanine ester 1-(2,3-dihydroxypropyl-1)-4-thiouracyl - AcPhePhe-HP-Ade and AcPheGly-HP-Ade acetyl-l-diphenylalanine and acetyl-l-phenylalanylglycine esters of HP-Ade respectively - AcPhe-puromycin acetyl-l-phenylalanyl-puromycin  相似文献   

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