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1.
Laran T. Jensen J. M. Peltier Dennis R. Winge 《Journal of biological inorganic chemistry》1998,3(6):627-631
Mammalian metallothioneins (MT) are known to maximally bind 12 copper ions in two six-Cu(I) ion clusters. Using electrospray
ionization mass spectrometry of MT at pH 4.5, a four-Cu(I) ion cluster was observed intermediate to a fully formed six Cu(I)
in a single domain or a fully formed Cu12MT species. The four-Cu(I) cluster was observed in both MT1 and MT3 isoforms. Addition of increasing amounts of Cu(I) to MT
at pH 4.5 resulted in prominent ions whoses masses were consistent with apo-MT, Cu4MT, Cu6MT, and Cu12MT. The cooperativity of cluster formation was reduced at pH 2.5. Addition of Cu(I) to apo-MT at a reduced pH resulted in
a series of ions consistent with Cu4 to Cu12MT species. However, formation of the tetracopper MT species remained cooperative at low pH, suggesting that this species
is very stable. To determine whether the tetracopper cluster was formed in either the α or β domain, domain peptides of MT3
were used. Addition of Cu(I) to the apo β domain resulted in a peak consistent with the formation of a four-Cu(I) cluster.
This is consistent with reports that Cu(I) ions bind preferentially to the β domain of MTs.
Received: 2 June 1998 / Accepted: 21 August 1998 相似文献
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4.
Yasufumi Kaneda Helene Hayes Tsuyoshi Uchida Michihiro C. Yoshida Yoshio Okada 《Chromosoma》1987,95(1):8-12
A human-mouse hybrid segregant HM76Dd40-6 with new characteristics was derived from the hybrid cell line HM76Dd containing human chromosome 19 as the only human chromosome. Three virus sensitivities located on human chromosome 19 (PVS, E11S and RDRC) were lost in HM76Dd40-6, while six other genes (C3, LDLR, EF2, GPI, PEPD and MANB) were retained. Cytogenetic analysis and in situ hybridization using human or mouse repeated sequences as probes showed that the region q13.1-qter of human chromosome 19 had been replaced by a fragment of mouse chromosome. Our results permit further regional assignment for the following five genes on human chromosome 19: GPI in the region cen-q12, MANB in p13.2-q12, E11S and RDRC in q13.1-qter, and EF2 in pter-q12. 相似文献
5.
The photorespiratory activity of mesophyll protoplasts of Nicotiana plumbaginifolia has been clearly demonstrated by the presence of a Warburg-effect, the occurrence of an important CO2-sensitive O2 uptake and the effect of some photorespiratory inhibitors on photosynthetic activity. At a nonsaturating dissolved inorganic carbon (DIC) concentration (0.1 millimolar), we observed that the rate of CO2 fixation was 60% lower at 50% O2 compared to that measured at 2% O2. Using 18O2 and mass spectrometry, we measured O2 exchange as a function of light intensity and of DIC concentration. Oxygen uptake measured at the CO2 compensation point (47.4 micromoles O2 per hour per milligram chlorophyll) was three-fold higher than that measured at a saturating CO2 concentration. Cyanide or iodoacetamide, inhibitors of the Calvin cycle, were found to reduce the O2 uptake to the same extent as CO2 saturation. We conclude from these results that the major part of the CO2-sensitive O2 uptake is due to photorespiration. Further, we investigated the effect on net photosynthesis of some inhibitors of the glycolate pathway. At CO2 saturation (10 millimolar DIC), 5 millimolar aminoacetonitrile (AAN), and 1 millimolar aminooxyacetate (AOA) did not cause any significant decrease in net photosynthesis. However, when these two inhibitors were added under a period of active photorespiration (10 minutes at the CO2 compensation point at 20% O2), we observed a decrease in the rate of net photosynthesis at 10 millimolar DIC measured afterward (respectively, 18 and 29%). This inhibition did not appear at 2% O2, but was stronger at 50% O2 (40% for AAN and 47% for AOA). With 0.05 millimolar butyl 2-hydroxy-3-butynoate (BHB) or 0.5 millimolar l-methionine-dl-sulfoximine (l-MSO), rates of net photosynthesis at 10 millimolar DIC were decreased by 10 to 15%. Additional decreases were observed after a period at the CO2 compensation point at 20% O2 (30% for BHB and 20% for l-MSO). From the sites of action of the four inhibitors tested, we suggest the inhibition of photosynthesis occurring after a period of active photorespiration to be due to the toxic accumulation of nonmetabolized phosphoglycolate. 相似文献
6.
Atrazine and diuron resistant clones were isolated from diploid photoautotrophic protoplastderived colonies of Nicotiana plumbaginifolia. Protoplasts were mutagenised with 0.1 mM N-ethyl-N-nitrosourea and colonies were screened for resistance after plating. Selection of calli was carried out on their ability to grow and green on a selective medium containing either atrazine or diuron. Plants were regenerated from most tolerant calli. Herbicide spray showed that plants of 6 and 4 clones were resistant to atrazine and diuron, respectively.Abbreviations Atrazine
2-chloro-4-ethylamino-6-isopropyl-amino-s-triazine
- diuron
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- NEU
N-ethyl-N-nitrosourea
- PSII
photosystem II 相似文献
7.
Two affinity states of M1 muscarine receptors 总被引:4,自引:0,他引:4
Lincoln T. Potter Cynthia A. Ferrendelli Helene E. Hanchett 《Cellular and molecular neurobiology》1988,8(2):181-191
1. The binding of oxotremorine-M to M1 muscarine receptors was examined by measuring competition between the agonist and 3H-pirenzepine, using rabbit hippocampal membranes suspended in 20 mM Tris buffer containing 1 mM Mn2+. 2. Both ligands interacted with a single class of receptors. The receptors could assume two affinity states for oxotremorine-M, with equal numbers of high-affinity (KH) and low-affinity (KL) sites. 3. KH interconverted reversibly to KL in the absence of divalent cations and interconverted reversibly to a state similar to KL in the presence of guanyl 5'-yl imidodiphosphate. 4. The results are compatible with a model in which a pair of receptor molecules can be stabilized by a guanine nucleotide-binding "G protein" and have one site each of KH and KL affinity. 相似文献
8.
Ammonia exchange and photorespiration in chlamydomonas 总被引:2,自引:2,他引:0
Two hours after the addition of l-methionine-dl-sulfoximine to the cell suspension, glutamine synthetase activity was inhibited by more than 90% in air-grown Chlamydomonas reinhardii. Cells continued to take up NH3 from the medium provided that the concentration of dissolved CO2 was high (equilibrated with 4% CO2 in air). This NH3 uptake, about 30% of the control, is discussed in terms of glutamate dehydrogenase activity. Without CO2, or with a low CO2 level, a NH3 excretion was observed, the rate of which depended on the actual concentration of the dissolved CO2. Experiments with 15NH3 demonstrated that no NH3 uptake was masked by this excretion and inversely that no excretion occurred during the uptake in the conditions where it took place. Furthermore, the NH3 excretion observed in the absence of CO2 increased when O2 concentration rose to 15% and was inhibited when 10 millimolar isonicotinic acid hydrazide was supplied to the algal suspension. Thus, NH3 excretion in the presence of l-methionine-dl-sulfoximine seems to be related to a photorespiratory process inasmuch as it presents the same properties with regard to the O2 and the isonicotinic acid hydrazide effects. These results favor the hypothesis that NH3 produced in the medium originates from the glycine to serine reaction. On the other hand, partial inhibition (50%) of photosynthesis by l-methionine-dl-sulfoximine was attributed to uncoupling between electron transfer and photophosphorylation due to NH3 accumulation into the cell. 相似文献
9.
Correlation Between Degradation of Bacteriophage T2 Deoxyribonucleic Acid and the Resistance of Escherichia coli to Infection 总被引:3,自引:2,他引:1
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The ability of certain strains of Escherichia coli to degrade T2 deoxyribonucleic acid to acid-soluble fragments is correlated with their high capacity to survive T2 infection. 相似文献
10.
Actions of Excitatory Amino Acids on Somatostatin Release from Cortical Neurons in Primary Cultures 总被引:1,自引:1,他引:0
L-Glutamate, N-methyl-D-aspartic acid (NMDA), quisqualate, and kainate were found to increase endogenous somatostatin release from primary cultures of rat cortical neurons in a dose-dependent manner. The rank order of potency calculated from the dose-response curves was quisqualate greater than glutamate = NMDA greater than kainate, with EC50 values of 0.4, 20, and 40 microM, respectively. Alanine, glutamine, and glycine did not modify the release of somatostatin. The stimulation of somatostatin release elicited by L-glutamate was Ca2+ dependent, was decreased by Mg2+, and was blocked by DL-amino-5-phosphonovaleric acid (APV) and thienylphencyclidine (TCP), two specific antagonists of NMDA receptors. The NMDA stimulatory effect was strongly inhibited by APV in a competitive manner (IC50 = 50 microM) and by TCP in a noncompetitive manner (IC50 = 90 nM). The release of somatostatin induced by the excitatory amino acid agonists was not blocked by tetrodotoxin (1 microM), a result suggesting that tetrodotoxin-sensitive, sodium-dependent action potentials are not involved in the effect. Somatostatin release in response to NMDA was potentiated by glycine, but the inhibitory strychnine-sensitive glycine receptor did not appear to be involved. Our data suggest that glutamate exerts its stimulatory action on somatostatin release essentially through an NMDA receptor subtype. 相似文献