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21,279 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A method is presented for obtaining the preparation of N+, K+-ATPase from the cattle brain. The specific activity of the preparation is 5 units (mu mole Pi per 1 min) per 1 mg of protein. A water-soluble derivate of carbodiimide is shown to inhibit reversibly both Na+, K+-ATPase and K+-phosphatase. ATP, Na+ and K+ manifest a protective effect against inhibition, and Na+ and K+ revealed a competition with the inhibitor for the enzyme. p-Chloromercuribenzoate inhibits irreversibly Na+, K+-ATPase and K+-phosphatase activities. The substrates ATP and p-nitrophenylphosphate protected these activities against inhibition. The phosphororganic compound O-n-butyl-S-(beta-ethyl-mercaptoethyl)-methyl thiophosphate has no significant effect on the Na+, K+-ATPase and K+-phosphatase activities.
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PMID:[Isolation of active preparation of Na+, K+-ATPase from cattle brain and study of the role of carboxyl, sulfhydryl and hydroxyl groups]. 12 66

The mechanical properties and the activity of the myofibrillar ATPase have been investigated at 21 degrees C on glycerinated back muscle from the water-bug Lethocerus colossicus. When the fibres were held under isometric conditions after stretching them by 0.5--4%, the ATPase required to maintain a given tension increases from 19 to 39 p-moles ATP split for each mg of tension developed as the Ca2+ level is increased from 10(-7) to up to 10(-5) M. The mechanical properties and the ATPase activity have been determined for Ca2+-activated fibres using sinusoidal frequencies of 1--30 HZ and oscillatory amplitudes of 0.5--6% peak-to-peak. In this way the R.M.S. velocity of sinusoidal movement was varied between 0.1-10 mm/sec. The rate of ATP splitting associated with oscillatory tension development, the dynamic tension cost, increases both with Ca2+ and with frequency of oscillation (at 1% peak-to-peak amplitude), becoming as high as four times the isometric value. The oscillatory power output which can be obtained is increased when the Ca2+ level is raised from 10(-7) to 10(-5) M or towards higher amplitudes of oscillation. The chemo-mechanical coupling efficiency increases proportionally with the R.M.S. velocity of muscle movement. In presence of 10(-5) M Ca2+ optimal efficiencies of 5.5--6.2 kcal work per mole ATP split are obtained at R.M.S. velocities of 1.3--2 muscle lengths/sec. The ability of the muscle fibres to perform osciillatory work at the higher frequencies was much reduced at lower Ca2+ levels of 10(-6) or 10(-7) M and the maximal efficiencies never exceeded 2.2 kcal/mole.
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PMID:The chemo-mechanical coupling relation in the oscillatory contraction-relaxation cycles of insect fibrillar muscle. 14 Feb 2

When zeaxanthin ([3R,3R']-beta, beta-carotene-3,3'diol) is inserted into phospholipid dispersions and the latter heated through their gel-liquid crystal phase transitions, large changes are noted in the resonance Raman and absorption spectra of the carotenoid molecule. By analogy with the data of Carey and co-workers (J. Raman Spectrosc. 6:282) who studied the aggregation of zeaxanthin in acetone-water solutions, it is suggested that the carotenoid aggregates in the phospholipid gel state while forming a monomer in liquid crystal phases. The alterations in both the visible absorption and resonance Raman data have been used to monitor phospholipid phase behavior in dipalmitoylphosphatidylcholine and distearoylphosphatidylcholine, (DSPC) one-component systems and binary mixtures. The phase diagram obtained for the binary system, as constructed from visible absorption and resonance Raman data, is compared with that of Shimshick and McConnell (Biochemistry. 12:2351) obtained from electron spin resonance (ESR) studies. Although the agreement between absorption and ESR data is generally satisfactory, onset temperatures for phase separation at low DSPC mole fractions deduced from resonance Raman measurements are several degrees lower than those from the other methods. Nevertheless, the use of zeaxanthin as a resonance Raman and visible absorption probe behavior will be useful in some situations where ordinary Raman spectroscopic data cannot be obtained easily. The advantage of the resonance Raman approach is illustrated in a study of the phase behavior of a phospholipid extract of a cel- mutant of Neurospora crassa. A phase separation region is observed with onset and completion temperatures of -19 and -6 degrees C, respectively.
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PMID:Zeaxanthin ([3R,3'R]-beta, beta-carotene-3-3'diol) as a resonance Raman and visible absorption probe of membrane structure. 16 48

Measurements of water proton spin relaxation enhancements (epsilon) can be used to discriminate high-affinity binding of Mn-2+ or Gd-3+ to biological membranes, from low-affinity binding. In rat liver mitochondria, epsilon b values of approx. 11 are observed upon binding of Mn-2+ to the inner membrane, while internal or low-affinity binding remains invisible to this technique. Energy-driven Mn-2+ uptake by liver mitochondria results in the subsequent decay of epsilon. Comparison of epsilon with the initial velocity of Mn-2+ uptake in rat liver mitochondria reveals a linear correlation, which holds at all temperatures between 0 degrees C and 40 degrees C, regardless of the mitochondrial protein concentration. Consequently, enhancement appears to reflect the binding of Mn-2+ to the divalent cation pump. Binding of Mn-2+ to blowfly flight muscle also results in substantial epsilon, which is associated with the glycerol-1-phosphate dehydrogenase instead of divalent cation transport. Consequently, no decay in epsilon due to uptake occurs after Mn-2+ is bound. Lanthanide ions are also bound and transported by mitochondria. Addition of Gd-3+ to pigeon heart or rat liver mitochondria results in epsilon b approximately equal to 5-6, which decays with similar kinetics in both systems. The uptake velocity of Gd-3+ in rat liver mitochondria is about 1/6 the rate with which Mn-2+ is transported. Lanthanides also diminish epsilon due to the addition of Mn-2+, and greatly retard the Mn-2+ uptake kinetics. The presence of carbonylcyanide-p-trifluoromethoxyphenylhydrazone depresses epsilon upon addition of Mn-2+ or Gd-3+ and also uncouples energy-driven uptake. On the other hand, prolonged anaerobic incubation in the presence of antimycin and rotenone exhausts the mitochondria of their energy stores, blocks the uptake of Mn-2+, but does not affect epsilon significantly. Evidently, the uncoupler-induced disappearance of divalent cation binding sites is not the result of "de-energization". Measurements of epsilon at several NMR frequencies indicate a correlation time (tau b) for carrier-bound Mn-2+ in rat liver mitochondria between 20 ns and 4 ns as one varies the temperature between 10 degrees C and 30 degrees C. The 13 Kcal/mole activation energy for tau b suggests that the 11 ns time constant at room temperature represents the movement of the Mn-11-carrier comples. On the other hand, tau b is probably approx. 100 times too short to represent the rotational motion of a carrier protein. Apparently, Mn-2+ binds to a small arm of the carrier which moves independent
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PMID:Magnetic resonance studies on the mitochondrial divalent cation carrier. 16 99

Oriented dipalmitoyllecithin-cholesterol multibilayers with 11% water have been studied with the cholestane spin label. From the ESR spectra the order parameters and the mobility of the spin label about its long axis have been calculated. The results on pure lecithin multibilayers indicate a transition from gel to liquid crystalline phase at 52 plus or minus 2 degrees C. In the gel phase the lecithin alkyl chains are highly ordered, but tilted with respect to the normal to the bilayers by about 25 degrees. Above 52 degrees C the tilt disappears and the mobility of the cholestane spin label increases, indicating an increase of mobility of the lecithin alkyl chains. When cholesterol is added, below about 52 degrees C a decrease of order is found. Furthermore, already small cholesterol contents (smaller than or equal to 10 mole %) remove the tilt. Above about 52 degrees C cholesterol improves the order by decreasing the amplitude of the librational motions. Cholesterol lowers the transition temperature of the system and reduces the mobility of the lecithin alkyl chains in the liquid crystalline phase. However an increase in mobility is found at cholesterol contents up to 10 mole %. A very broad phase transition is observed at 50 mole % cholesterol. In all systems an increase in temperature results in a reduction of order through an increase of the amplitude of the librational motions of the molecules. The librational motions are to some extent cooperative. The asymmetry of the order matrix is found to be a measure for the lateral ordering. Cholesterol increases the lateral ordering, indicating that the flat cholesterol molecules orient parallel to each other.
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PMID:An ESR Spin label study of structural and dynamical properties of oriented lecithin-cholesterol multibilayers. 16 12

Human serum lipoproteins and egg yolk lecithin liposomes are able to solubilize large amounts of n-hexane and n-octane. At the maximum water solubility of n-octane the mole ratio of alkane to lipoprotein was 65 for high density lipoprotein (holo-HDL) and 900 for low density lipoprotein (holo-LDL). Alkane binding to lipid-free apo-HDL is negligible compared to alkane solubility in holo-HDL. Alkane solubility in the lipoproteins and liposomes is thermodynamically consistent with the simple soution of hydrocarbon in the hydrophobic regions of these particles. The unitary free energies of alkane transfer are similar to values previously observed for detergent micelles but are less favorable by 0.8 kcal/mol from the free energy of transfer to liquid hydrocarbon. It is concluded that the thermodynamics of alkane transfer to the lipoproteins resembles that found for detergent micelles or liposomes rather than that anticipated for an "oil drop" (i.e. liquid hydrocarbon).
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PMID:Hydrophobic interaction of alkanes with liposomes and lipoproteins. 16 99

2H and 31P spin-lattice relaxation times (T1) were studied for inverted egg phosphatidylcholine micelles in CCl4 as functions of 2H2O concentration. When the 2h2O/phosphatidylcholine mole ratio changed from 1.0 to 18.0, T1 of 31P increased by about 2.6 fold, whereas T1 of 2H increased by about 50 fold. A quantitative analysis of the deuterium T1 data showed that there is only one water molecule tightly bound to the polar head, and it is in rapid exchange with the rest of the water molecules. The activation energy for the deuterium T1 was 7.1 +/- 0.8 kcal/mol(30 +/- 3 kJ/mol), and was independent of the 2H2O concentration.
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PMID:The interaction between water and the polar head in inverted phosphatidylcholine micelles. A 2H and 31P relaxation study. 18 57

We used paired-ion high-performance liquid chromatography to determine the 4-nitrophenol content of 4-nitrophenyl phosphate, a substrate for alkaline phosphatase analysis. This was done on a reversed-phase column with a mobile phase of methanol/water, 45/55 by vol, containing 3 ml of tetrabutylammonium phosphate reagent per 200 ml of solvent. At a flow rate of 1 ml/min, 4-nitrophenol was eluted at 9 min and monitored at 404 nm; 4-nitrophenyl phosphate was eluted at 5 min and could be monitored at 311 nm. Samples of 4-nitrophenyl phosphate obtained from several sources contained 0.3 to 7.8 mole of 4-nitrophenol per mole of 4-nitrophenyl phosphate.
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PMID:4-Nitrophenol in 4-nitrophenyl phosphate, a substrate for alkaline phosphatase, as measured by paired-ion high-performance liquid chromatography. 20 Mar 79

1. The amounts of endogenous serotonin (5-HT) released into the medium by the cerebro-buccal ganglionic ring of Aplysia californica incubated in artificial sea water (ASW) were measured. The rate of spontaneous 5-HT release varied between 0.4 and 1.2 p-mole per hour, which is less than 1% of the total 5-HT present in this preparation.2. Direct stimulation of the ordinarily silent 5-HT-containing giant cerebral neurones resulted in a 80-100% increase of the 5-HT released, but only when the 5-HT uptake was blocked by chlorimipramine (1-10 muM).3. High K(+) media (50 mM) also caused a significant increase in the amount of 5-HT released from the preparation provided that chlorimipramine (1-10 muM) was present in the incubation fluid.4. Co(2+) ions (10-30 mM) added to the incubating medium blocked the spontaneous leak of endogenous 5-HT as well as the release, in the presence of chlorimipramine, evoked either by stimulation of the 5-HT-giant cerebral neurones or high K(+)-media.5. In the presence of chlorimipramine or desmethylimipramine, the duration and/or the amplitude of the excitatory or the inhibitory synaptic potentials evoked in the buccal neurones by the stimulation of the 5-HT giant cerebral neurones were markedly enhanced.6. These results strongly support the idea that 5-HT is the synaptic transmitter released at the excitatory and inhibitory junctions established by the 5-HT giant cerebral neurones in the ipsilateral buccal ganglia. In addition, they underline the role of amine re-uptake in the physiological inactivation of 5-HT as a transmitter.
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PMID:Release of endogenous serotonin from two identified serotonin-containing neurones and the physiological role of serotonin re-uptake. 20 73

1. The chloride equilibrium flux (chloride self-exchange) was determined by measuring the rate of 36Cl efflux from radioactively labelled human red cells. The cellular chloride concentration was varied between 5 and 700 mM by the nystatin technique (Cass & Dalmark, 1973). The chloride transport capacity was not affected by the nystatin technique. 2. The chloride equilibrium flux showed saturation kinetics in the pH range between 6-2 and 9-2 (0 degrees C). The chloride transport decreased at chloride concentrations higher than those which gave the maximum transport. 3. The apparent half-saturation constant, (K1/2), depended on the pH and whether the chloride transport was perceived as a function of the chloride concentration in the medium or in the cell water. The (K1/2)m increased and the (K1/2)c decreased with increasing pH. The dependence of the chloride transport on the chloride concentration was described by Michaelis-Menten kinetics at pH 7-2, but at values of pH outside pH 7-8 S-shaped or steeper graphs were observed. 4. The chloride equilibrium flux varied with the pH at constant chloride concentration in the medium (pH 5-7-9-5). The transport had a bell-shaped pH dependence at chloride concentrations below 200 mM. At chloride concentrations between 300 and 600 mM the chloride transport increased with increasing pH to reach a plateau around pH 8. The position of the acidic branches of the pH graphs was independent of the chloride concentration (25-600 mM), but the position of the alkaline branches moved towards higher values of pH with increasing chloride concentration (5-150 mM). Thus, the position of the pH optimum increased with increasing chloride concentration. The chloride transport at low pH values was a function of the inverse second power of the hydrogen ion concentration. The pK of the groups which caused the inhibition was approximately 6 and independent of the temperature (0-18 degrees C). 5. The chloride equilibrium flux as a function of chloride concentration, pH, and temperature could be described by a transport model with a mobile, positively charged, chloride binding carrier with a single chloride dissociation constant of 33 mM, a transport capacity of 900 m-mole/3 x 10(13) cells.min (pH 7-2, 0 degrees C), and an Arrhenius activation energy of 30 kcal/mole. The pH dependence of the transport of inorganic monovalent and divalent anions is discussed in relation to the suggested model.
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PMID:Chloride transport in human red cells. 24 Sep 29


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