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11.
Blue bacteriorhodopsin was prepared by electrodialysis, cation-exchange chromatography and acidification. The electrooptical properties of these preparations compared to those of the native purple bacteriorhodopsin suggest that the blue bacteriorhodopsin has a smaller induced dipole moment than the native purple bacteriorhodopsin and that bound cations in the native bacteriorhodopsin stabilize the protein conformation in the membrane.Purple bacteriorhodopsin was regenerated by addition of potassium, magnesium or ferric ions to blue bacteriorhodopsin. Both spectrscopically and electrooptically the potassium- and ferric-regenerated samples are different from the native purple state. Although the magnesium-regenerated sample is spectroscopically similar to the native purple bacteriorhodopsin, the electrooptical properties are rather similar to those of the cation-depleted blue sample, suggesting that it is very difficult to re-stabilize protein structures once cations are depleted. 相似文献
12.
Scavenging of superoxide radical by ascorbic acid 总被引:1,自引:0,他引:1
Using acetaldehyde and xanthine oxidase as the source of suPeroxide radical, the second order rate constant for the reaction
between ascorbic acid and superoxide radical was estimated to be 8.2 X 107 M-1 s-1. In rats, the average tissue concentration of ascorbic acid was of the order of 10-3 M and that of superoxide dismutase was of the order of 10-6 M. So, taking together both the rate constants and the tissue concentrations, the efficacy of ascorbic acid for scavenging
superoxide radical in animal tissues appears to be better than that of suPeroxide dismutase. The significance of ascorbic
acid as a scavenger of superoxide radical has been discussed from the point of view of the evolution of ascorbic acid synthesizing
capacity of terrestrial vertebrates. 相似文献
13.
Glutamine synthetase (GS) from the chick brain was purified to apparent homogeneity by ammonium sulfate fractionation followed by affinity chromatography, electrofocusing and Sephadex G-150 chromatography. The purified enzyme showed a single band on sodium dodecyl sulfate analysis in polyacrylamide gel. By sedimentation equilibrium analysis and gel electrophoresis analysis, it was shown that the enzyme has a subunit molecular weight of 45,000 and a native molecular weight of 364,000, which is consistent with an octameric structure. Sedimentation analysis in the presence of Mg2+ revealed three different forms of macromolecules corresponding respectively to a monomer, a tetramer and an octamer. Among eight cations tested (Ca2+, Co2+, Fe2+, Li+, Mg2+, Mn2+, Ni2+, Zn2+) only Co2+, Mg2+ and Mn2+ supported GS activity; the order of activatory ability was Mg2+>Co2+>Mn2+. The maximum activating effect of Mn2+ occurs only within a very narrow range of concentration: with an excess of cation causing strong inhibition of GS activity. For each cation, maximal GS activity occurs at a defined cation/ATP ratio. A regulatory system in which Mn2+, modulates the Mg2+ dependent GS activity, is proposed; such cation interactions may be of significance in the intracellular control of glutamine synthesis. 相似文献
14.
Curtis W. Hoganson Demetrios F. Ghanotakis Gerald T. Babcock Charles F. Yocum 《Photosynthesis research》1989,22(3):285-293
Manganese in the oxygen-evolving complex is a physiological electron donor to Photosystem II. PS II depleted of manganese may oxidize exogenous reductants including benzidine and Mn2+. Using flash photolysis with electron spin resonance detection, we examined the room-temperature reaction kinetics of these reductants with Yz
+, the tyrosine radical formed in PS II membranes under illumination. Kinetics were measured with membranes that did or did not contain the 33 kDa extrinsic polypeptide of PS II, whose presence had no effect on the reaction kinetics with either reductant. The rate of Yz
+ reduction by benzidine was a linear function of benzidine concentration. The rate of Yz
+ reduction by Mn2+ at pH 6 increased linearly at low Mn2+ concentrations and reached a maximum at the Mn2+ concentrations equal to several times the reaction center concentration. The rate was inhibited by K+, Ca2+ and Mg2+. These data are described by a model in which negative charge on the membrane causes a local increase in the cation concentration. The rate of Yz
+ reduction at pH 7.5 was biphasic with a fast 400 s phase that suggests binding of Mn2+ near Yz
+ at a site that may be one of the native manganese binding sites.Abbreviations PS II
Photosystem II
- YD
tyrosine residue in Photosystem II that gives rise to the stable Signal II EPR spectrum
- Yz
tyrosine residue in Photosystem II that mediates electron transfer between the reaction center chlorophyll and the site of water oxidation
- ESR
electron spin resonance
- DPC
diphenylcarbazide
- DCIP
dichlorophenolindophenol 相似文献
15.
Kenneth S. Leonards 《Molecular and cellular biochemistry》1990,95(1):31-42
To ascertain the roles of the membrane proteins in cation/sarcolemmal membrane binding, isolated rat cardiac sarcolemmal vesicles were extensively treated with Protease (S. aureus strain V.8). SDS-gel electrophoresis, protein and phosphate analysis confirmed that at least 20–22% of the protein, but none of the phospholipid, was solubilized by this procedure, and that the remaining membrane proteins were extensively hydrolyzed into small fragments. The cation binding properties of the treated vesicles were then examined by analyzing their aggregation behavior. The results demonstrate that this procedure had no effect on the selectivity series for di- and trivalent cation binding, or the divalent cation-induced aggregation behavior of the sarcolemmal vesicles at different pHs, indicating that proteins are probably not involved in these interactions and cannot be the low affinity cation binding sites previously observed [21, 22]. It did, however, change the pH at which protons induced sarcolemmal vesicle aggregation, suggesting a possible role for proteins in these processes. Protease treatment also modified the effects of fluorescamine labelling on divalent cation-induced vesicle aggregation, indicating that the NH, groups being labelled with fluorescamine are located on the sarcolemmal proteins. Together, these results support the hypothesis that di- and trivalent cation binding to the sarcolemmal membrane is largely determined by lipid/lipid and/or lipid/carbohydrate interactions within the plane of the sarcolemmal membrane, and that membrane proteins may exert an influence on these interactions, but only under very specialized conditions.Abbreviations MES
2-(N-morpholino)ethanesulfonic acid
- MOPS
3-(N-morpholino) propanesulfonic acid
- HEPES
N-2-Hydroxyethylpiperizine-N-2- ethanesulfonic acid
- CHES
2(N-Cyclohexylamino) ethanesulfonic acid
- DTT
DL-Dithiothreitol
- PMSF
Phenylmethyl-sulfonyl fluoride 相似文献
16.
Calcium ion transport across discs of the cortical flesh of apple fruit in relation to fruit development 总被引:3,自引:0,他引:3
Transport of Ca2+ through discs of apple fruit tissue was examined in tissue taken at different stages of fruit development. Transport rates decreased with fruit development when cation exchange was the predominant influence on transport (with 10−6 M 45 CaCl2 as the source solution). This decrease was associated with a reduction in relative cell wall surface area, cation exchange capacity and cell wall yield that occurred during fruit growth. When diffusion was the major transport force, and when transport was influenced by solution infiltration of the tissue disc (10−2 M 45 CaCl2 in the source solution), transport rates increased during fruit growth. This increment was related to increases in air space of the tissue. Ca2+ transport through apple fruit tissue is influenced by the extent and nature of the cell wall, changing proportions of air space and Ca2+ concentration in the extracellular solution. 相似文献
17.
Roberta Colombo Raffaella Cerana Piera Lado Antonio Peres 《The Journal of membrane biology》1988,103(3):227-236
Summary The patch-clamp technique in whole-cell configuration was used to study the electrical properties of the tonoplast in isolated vacuoles fromAcer pseudoplatanus cultured cells. In symmetrical KCl or K2 malate solutions, voltage- and time-dependent inward currents were elicited by hyperpolarizing the tonoplast (inside negative), while in the positive range of potential the conductance was very small. The specific conductance of the tonoplast at –100 mV, in 100mm symmetrical KCl was about 160 S/cm2. The reversal potentials (E
rev) of the current, measured in symmetrical or asymmetrical ion concentrations (cation, anion or both) were very close to the values of the K+ equilibrium potential. Experiments performed in symmetrical or asymmetrical NaCl indicate that Na+ too can flow through the channels. NeitherE
rev nor amplitude and kinetics of the current changed by replacing NaCl with KCl in the external solution. These results indicate the presence of hyperpolarization-activated channels in tonoplasts, which are permeable to K+ as well as to Na+. Anions such as Cl– or malate seem to contribute little to the channel current. 相似文献
18.
Richard C. Woodman John T. Curnutte Bernard M. Babior 《Free radical biology & medicine》1988,5(5-6):355-361
We examined the effects of the recombinant human colony stimulating factors GM-CSF and G-CSF, cycloheximide (a protein synthesis inhibitor) and dihydrocytochalasin B (a microfilament disrupting agent) upon FMLP (N-formyl-methionyl-leucylphenylalanine)-stimulated O2 − production by neutrophils. We confirmed a time dependent augmentation of O2 − production following preincubation of neutrophils either alone or with colony stimulating factors. Furthermore, we found that GM-CSF, but not G-CSF, increased O2 − production at some concentrations of the stimulus. Preincubation of neutrophils with cycloheximide in the absence of CSF caused a marked fall in O2−-production that was first evident at 2 hours. The fall in O2−-forming capacity caused by cycloheximide was much less pronounced if dihydrocytochalasin B was also included in the preincubation buffer. These findings suggest a previously unrecognized role for de novo protein synthesis in maintaining the ability of neutrophils to manufacture O2−, and support earlier studies indicating that the cycling of FMLP receptors between the cell membrane and an intracellular compartment is important in determining the magnitude of the respiratory burst in FMLP-stimulated neutrophils. 相似文献
19.
Robert E. Beyer 《Free radical biology & medicine》1988,5(5-6):297-303
The ability of coenzyme Q to inhibit lipid peroxidation in intact animals as well as in mitochondrial, submitochondrial, and microsomal systems has been tested. Rats fed coenzyme Q prior to being treated with carbon tetrachloride or while being treated with ethanol excrete less thiobarbituric acid-reacting material in the urine than such rats not fed coenzyme Q. Liver homogenates, mitochondria, and microsomes isolated from rats treated with carbon tetrachloride and ethanol catalyze lipid peroxidation at rates which exceed those from animals also fed coenzyme Q. The rate of lipid peroxidation catalyzed by submitochondrial particles isolated from hearts of young, old, and endurance trained elderly rats was inversely proportional to the coenzyme Q content of the submitochondrial preparation in assays in which succinate was employed to reduce the endogenous coenzyme Q. Reduced, but not oxidized, coenzyme Q inhibited lipid peroxidation catalyzed by rat liver microsomal preparations. These results provide additional evidence in support of an antioxidant role for coenzyme Q. 相似文献
20.
Klaas J. Lusthof Nicolaas J. del Mol Wilma Richter Lambert H.M. Janssen John Butler Brigid M. Hoey Willem Verboom David N. Reinhoudt 《Free radical biology & medicine》1992,13(6):599-608
The formation of reactive oxygen intermediates (ROI) during redox cycling of newly synthesized potential antitumor 2,5-bis (1-aziridinyl)-1,4-benzoquinone (BABQ) derivatives has been studied by assaying the production of ROI (superoxide, hydroxyl radical, and hydrogen peroxide) by xanthine oxidase in the presence of BABQ derivatives. At low concentrations (< 10 microM) some BABQ derivatives turned out to inhibit the production of superoxide and hydroxyl radicals by xanthine oxidase, while the effect on the xanthine-oxidase-induced production of hydrogen peroxide was much less pronounced. Induction of DNA strand breaks by reactive oxygen species generated by xanthine oxidase was also inhibited by BABQ derivatives. The DNA damage was comparable to the amount of hydroxyl radicals produced. The inhibiting effect on hydroxyl radical production can be explained as a consequence of the lowered level of superoxide, which disrupts the Haber-Weiss reaction sequence. The inhibitory effect of BABQ derivatives on superoxide formation correlated with their one-electron reduction potentials: BABQ derivatives with a high reduction potential scavenge superoxide anion radicals produced by xanthine oxidase, leading to reduced BABQ species and production of hydrogen peroxide from reoxidation of reduced BABQ. This study, using a unique series of BABQ derivatives with an extended range of reduction potentials, demonstrates that the formation of superoxide and hydroxyl radicals by bioreductively activated antitumor quinones can in principle be uncoupled from alkylating activity. 相似文献