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81.
The 1H-NMR spectrum of cucumber basic blue protein (CBP) has been recorded. Examination of the spectrum of the reduced protein suggests that one or more sidechains exist in conformations which interconvert slowly at ambient temperatures. His 39, His 84 and Met 89 are identified as copper ligands by redox titration and by amino acid sequence homology with plastocyanin and azurin. The importance of a Phe sidechain close to the Met ligand in the potential blue copper site is confirmed. Broadening of His ligand resonances at elevated temperatures reveals an exchange process at the reduced copper centre. 相似文献
82.
Initial Cu++ sorption by Cyclotella meneghiniana Kütz. (Cu++-sensitive) and Chlamydomonas reinhardtii Dangeard (Cu++-resistant) was rapid in the first 5 min of Cu++ incubation with little sorption after 2 h. On a cell to cell basis, Cyclotella sorbed ca. five times more Cu++ from the medium than Chlamydomonas. In MBL medium with EDTA Cyclotella and Chlamydomonas cells sorbed 21.0 and 4.41 nM Cu++/106 cells respectively in 6 h with 0.3 mg Cu++/l in the medium. Proportionally similar quantities of Cu++ were sorbed when the cells were Cu++ incubated in MBL + citrate or filtered lake water. Cleaned cell walls of Cyclotella sorbed little Cu++ (1.7 nM/106 cells) as compared to living cells (17.5 nM Cu++/106 cells) in 3 h. Therefore, in living Cyclotella most of the Cu++ taken up must be absorbed by the protoplasm or perhaps by the organic layer surrounding the silica wall. Cleaned cell walls of Chlamydomonas sorbed 3.5 nM Cu++/106 cells and living Chlamydomonas cells sorbed 2.6 nM Cu++/106 cells. This indicates that most of the Cu++ sorbed by Chlamydomonas cells remained bound to the cell wall and probably did not readily enter into the protoplasm: When placed in Cu++ free medium after Cu++ incubation, Cyclotella and Chlamydomonas cells released 46 and 59% respectively of the Cu++ sorbed. 相似文献
83.
Carl L. Keen Bo Lönnerdal Theodore N. Stein Lucille S. Hurley 《Biological trace element research》1980,2(3):221-227
The presence of superoxide dismutase in bovine and human milk was investigated by ultrafiltration, gel filtration, and isoelectric
focusing. Conclusive evidence for the presence of this enzyme in both milks is presented. The molecular weight of the enzyme
was estimated by gel filtration on Sephadex G-100 to be 30,000, which is consistent with reported values for the copper, zinc
form of superoxide dismutase. In addition, enzyme activity was inhibited by cyanide, thus eliminating the possibility that
the enzyme was present in the manganese form. Several isoenzymes were detected by isoelectric focusing in polyacrylamide gel,
and the isoenzyme pattern in bovine milk was the same as that found for bovine plasma, suggesting that milk superoxide dismutase
originates from plasma. It may be that the presence of copper, zinc superoxide dismutase in milk is important for the maintenance
of its oxidative stability. 相似文献
84.
Activities of Phosphorylase, glyceraldehyde-3 -phosphate dehydrogenase, lactate dehydrogenase, malate dehydrogenase and succinate
dehydrogenase in the rat endometrial tissue are significantly inhibited by an intrauterine copper device, while it stimulated
glucose-6-phosphate dehydrogenase activity. The copper device decreased the lactate/pyruvate ratio in the tissue; pyruvate
utilizationin vitro by the rat endometrium is also blocked by copper. These findings suggested that the normal carbohydrate metabolism of the
tissue may be affected in presence of copper, thus resulting in a change of the endometrial function, which may be one of
the factors responsible for the contraceptive and pharmacological action of an intrauterine copper device. 相似文献
85.
Andrzej J. Żelazowski Jadwiga A. Szymańska Czesklaw S. Cierniewski 《Chemico-biological interactions》1980,33(1):115-125
Two homogenous fractions of hepatic metallothioneins ((Cd,Zn) MT-1 and (Cd,Zn) MT-2) and renal metal binding proteins ((Bi,Cu) BP-1 and (Bi,Cu) BP-2) were isolated from rats exposed to heavy metals and specific antisera to them were produced in rabbits.These antisera were tested by immunodiffusion and immunoelectrophoresis for their ability to bind different fractions of hepatic Cd,Zn -metallothionein and renal (Bi,Cu)-, (Hg,Cu)- and (Cd,Cu)-binding proteins. It was found that anti (Bi,Cu) BP antisera did not cross-react with hepatic (Cd,Zn) MT-1 and (Cd,Zn) MT-2. Strong immunological cross-reactions were detected between anti (Bi,Cu) BP antisera and individual forms of (Cd,Cu)-, (Hg,Cu)- and (Bi,Cu)-binding proteins isolated from rat kidneys. 相似文献
86.
The stability of the serine proteases from Bacillus amyloliquefaciens (subtillisin BPN') and Bacillus licheniformis (subtilisin Carlsberg) was investigated in various anhydrous solvents at 45 degrees C. The half-life of subtilisin BPN' in dimethyl-formamide dramatically depends on the pH of the aqueous solutions from which the enzyme was lyophilized, increasing from 48 min to 20 h when the pH is raised from 6.0 to 7.9. Both subtilisins exhibited substantial inactivation during multihour incubations in tert-amyl alcohol and acetonitrile when enzymatic activities were also measured in these solvents; however, when the enzymes were assayed in water instead, hardly any loss of activity was detected. This surprising difference appears to stem from the partitioning of the bound water essential for catalytic activity from the enzymes into the solvents. When assayed in organic solvents, this time-dependent stripping of water results in decay of enzymatic activity; however, when assayed in water, where the dehydrated subtilisins can undergo rehydration thereby recovering catalytic activity, little inactivation is observed. In agreement with this hypothesis, the addition of small quantities of water tert-amyl alcohol stabilized the subtilisins in it even when enzymatic activity was measured in the nonaqueous solvent. Ester substrates (vinyl butyrate and trichloroethyl butyrate) greatly enhanced the stability of both subtilisins in organic solvents possibly because of the formation of the acyl-enzymes. 相似文献
87.
Washed or growing E. coli cells are killed by epinephrine, norepinephrine or dopamine in the presence of non lethal concentrations of Cu(II). Killing is enhanced by anoxia and by sublethal Concentrations of H2O1. The rate of killing is proportional to the rate of catecholamine oxidation. The copper epinephrine complex binds to E. coli cells, induces membrane damage and depletion of the cellular ATP pool. The cells may be partially protected by SOD or catalase but not by OH radical scavengers. Addition of H2O2 to cells which were sensitized by preincubation with the epinephrine-copper complex, causes rapid killing and DNA degradation. Sensitized cells are not protected by BSA. 相似文献
88.
Undesirable aggregation of aqueous insulin solutions remains a serious obstacle in the development of alternative methods of diabetes therapy. We investigated the fundamental nature of the aggregation mechanism and proposed stabilization strategies based on a mathematical model for the reaction scheme. Insulin aggregation kinetics in the presence of solid-liquid and air-liquid interfaces were monitored using UV spectroscopy and quasielastic light scattering (QELS). Experimental observations were consistent with our model of monomer denaturation at hydrophobic surfaces followed by the formation of stable intermediate species which facilitated subsequent macroaggregation. The model was used to predict qualitative trends in insulin aggregation behavior, to propose stabilization strategies, and to elucidate mechanisms of stabilization. In the absence of additives, insulin solutions aggregated completely (more than 95% of the soluble protein lost) within 24 h; with sugarbased nonionic detergents, no detectable loss occurred for more than 6 weeks. (c) 1992 John Wiley & Sons, Inc. 相似文献
89.
A. M. Rofe J. C. Philcox D. R. Haynes M. W. Whitehouse P. Coyle 《Biological trace element research》1992,34(3):237-248
The early changes in hepatic metallothionein (MT) and plasma zinc (Zn), copper (Cu), and iron (Fe) were investigated during
the induction of adjuvant (AJ) arthritis in rats in conjunction with cyclosporin (CSA) treatment. Plasma Zn decreased after
AJ injection (60% of control values at 8 h), and this was associated with a 4.5-fold increase in hepatic MT at 8 h. Plasma
Zn was lowest at 16 h (40% of control), whereas hepatic MT concentrations increased to a maximum of 20-fold at 16 h. Changes
in plasma Fe paralleled those of Zn, whereas plasma Cu levels were increased. Plasma metal and hepatic MT concentrations returned
toward normal from d 1–7. At d 14, when marked paw swelling was apparent, hepatic MT and plasma Cu were again increased and
plasma Zn decreased.
Administration of CsA decreased MT induction in rats injected with AJ and also caused a marked recovery in plasma Zn and Fe
levels. These changes were small but significant even in the early stages (up to 24 h) after AJ injection and were followed
by a sustained improvement in all parameters, corresponding to the nonappearance of clinical arthropathy in CsA-treated rats.
TNF-α and IL-6 production by peritoneal macrophages isolated from AJ-injected rats was significantly decreased by CsA treatment
at d 7 and 14. The inhibition of hepatic MT induction during acute and chronic inflammation by cyclosporin emphasizes the
role of the immune system in altered metal homeostasis in inflammation. 相似文献
90.
Thyroid hormones are involved in copper and zinc distribution in rat tissues. We examined the influence of thyroparathyroidectomy
(TPTY) and of a replacement therapy by T4 on Cu and Zn organ distribution. MT levels were also measured both in basal conditions
and after induction by cadmium. The results confirm that a lack of T4 modified Cu and Zn in serum and tissues. In serum, TPTY
increased Cu (+15%) and ceruloplasmin (+18%), and decreased Zn (−18%). In tissues, Cu was altered in liver (+13%), kidney
(−24%), heart (−16%) duodenum (−18%), and Zn in liver (+25%) and kidney (−10%). The soluble fractions (100,000 g supernatant)
were mainly affected in liver and kidney, and the subcellular fractions in heart and duodenum. MT levels were modified in
basal conditions only in liver (+57%) and kidney (−36%). T4 administration partially prevented the effect of TPTY on both
elements and MT concentrations. Therefore, no evidence is provided for a direct role of T4 in the metabolism of MT in a way
comparable to the effects of glucocorticoids. However, MT could mediate the consequences of TPTY on metal distribution in
certain organs, such as liver and kidney. 相似文献