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Summary Equivalent-circuit impedance analysis experiments were performed on the urinary bladders of freshwater turtles in order to quantify membrane ionic conductances and areas, and to investigate how changes in these parameters are associated with changes in the rate of proton secretion in this tissue. In all experiments, sodium reabsorption was inhibited thereby unmasking the electrogenic proton secretion process. We report the following: (1) transepithelial impedance is represented exceptionally well by a simple equivalent-circuit model, which results in estimates of the apical and basolateral membrane ionic conductances and capacitances; (2) when sodium transport is inhibited with mucosal amiloride and serosal ouabain, the apical and basolateral membrane conductances and capacitances exhibit a continual decline with time; (3) this decline in the membrane parameters is most likely caused by subtle time-dependent changes in cell volume, resulting in changes in the areas of the apical and basolateral membranes; (4) stable membrane parameters are obtained if the tissue is not treated with ouabain, and if the oncotic pressure of the serosal solution is increased by the addition of 2% albumin; (5) inhibition of proton secretion using acetazolamide in CO2 and HCO
3
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-free bathing solutions results in a decrease in the area of the apical membrane, with no significant change in its specific conductance; (6) stimulation of proton transport with CO2 and HCO
3
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-containing serosal solution results in an increase in the apical membrane area and specific conductance. These results show that our methods can be used to measure changes in the membrane electrophysiological parameters that are related to changes in the rate of proton transport. Notably, they can be used to quantify in the live tissue, changes in membrane area resulting from changes in the net rates of endocytosis and exocytosis which are postulated to be intimately involved in the regulation of proton transport. 相似文献
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Further characterization of type 2 and type 3 chain blood group A glycosphingolipids from human erythrocyte membranes 总被引:3,自引:0,他引:3
Blood group A glycosphingolipids with slow chromatographic mobilities have been separated systematically with an improved chromatographic procedure, and their structures have been analyzed by application of a panel of monoclonal antibodies defining A determinants carried by type 1, type 2, type 3, and type 4 carbohydrate chains as well as by 1H NMR spectroscopy and methylation analysis. Of several A-active fractions, previously termed Aa, Ab, Ac, and Ad, in decreasing order of thin-layer chromatographic mobility, the third fraction (Ac) was characterized as containing one type 3 chain A component and one type 2 chain A component without branching, which have been termed type 3 chain Ab and nor-Ac, respectively. (Formula: see text). The major component present in the fourth A-active fraction (Ad) was isolated and characterized as a branched type 2 chain glycolipid formerly termed Ac. The major component in the fifth A-active fraction (Ae) was identified as a branched type 2 chain A previously termed Ad. The structures of Ac (n = 1) and Ad (n = 2) are (Formula: see text). 相似文献
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In two groups of young healthy subjects who performed arm training (N = 5) and leg training (N = 5), respectively, the respiratory adaptation to submaximal exercise with trained and nontrained muscle groups was compared by measurement of the ventilatory equivalent (Ve/Vo2, pH, and blood gases (Pco2, Po2, and So2) in arterial blood and in venous blood from exercising extremities. After training Ve/Vo2 was significantly reduced during exercise with trained muscles, but unchanged during exercise with nontrained muscles. The reduction in Ve/Vo2 was closely related to a less pronounced increase in heart rate and in arterial lactate content, but showed no quantitative correlation to changes in arterial adaptations in trained muscles are mainly responsible for the reduction in Ve/Vo2. After training during exercise with trained as well as nontrained muscles a shift to the right of the blood oxygen dissociation curve occurred as extremities was lower while corresponding Po2 was higher. 相似文献
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Torben Clausen Tove Lindahl Andersen Marianne Stürup-Johansen Olga Petkova 《生物化学与生物物理学报:生物膜》1981,646(2):261-267
(1) The effects of vanadate of hexose transport, 45Ca-exchange and (Na+, K+)-contents have been characterized in isolated adipose tissue and skeletal muscles of the rat. (2) In whole epididymal fat pads, vanadate (0.5–5.0 mM) markedly stimulated the uptake of 2-deoxyl[14C]glucose as well as the efflux of . (3) Within the same concentration range, vanadate induced an early increase in 45Ca-washout from preloaded fat pads. The maximum increases in the fractional losses of and 45Ca were significantly correlated (, ). (4) In extensor digitorum longus and soleus muscles, vanadate (0.5–5.0 mM) stimulated the efflux of and this effect was preceded by a rise in the washout of 45Ca. The maximum increases in the fractional losses of and 45Ca were significantly correlated (, ). (5) In extensor digitorum longus and soleus muscles, vanadate increased K+-contents and decreased Na+ contents. (6) The stimulation of 45Ca-washout presumably reflects an increase in the cytoplasmic Ca2+ level, brought about by an inhibitory effect of vanadate on the Ca2+-sensitive ATPase of the sarcoplasmic or the endoplasmic reticulum. As demonstrated for most other insulin-like agents (Sørensen, S.S., Christensen, F. and Clausen, T. (1980) Biochim. Biophys. Acta 602, 433–445), the stimulating effect of vanadate on glucose transport appears to be associated with or mediated by a rise in the cytoplasmic Ca2+ level. 相似文献
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C Steegborn A Messerschmidt B Laber W Streber R Huber T Clausen 《Journal of molecular biology》1999,290(5):983-996
Cystathionine gamma-synthase catalyses the committed step of de novo methionine biosynthesis in micro-organisms and plants, making the enzyme an attractive target for the design of new antibiotics and herbicides. The crystal structure of cystathionine gamma-synthase from Nicotiana tabacum has been solved by Patterson search techniques using the structure of Escherichia coli cystathionine gamma-synthase. The model was refined at 2.9 A resolution to a crystallographic R -factor of 20.1 % (Rfree25.0 %). The physiological substrates of the enzyme, L-homoserine phosphate and L-cysteine, were modelled into the unliganded structure. These complexes support the proposed ping-pong mechanism for catalysis and illustrate the dissimilar substrate specificities of bacterial and plant cystathionine gamma-synthases on a molecular level. The main difference arises from the binding modes of the distal substrate groups (O -acetyl/succinyl versusO -phosphate). Central in fixing the distal phosphate of the plant CGS substrate is an exposed lysine residue that is strictly conserved in plant cystathionine gamma-synthases whereas bacterial enzymes carry a glycine residue at this position. General insight regarding the reaction specificity of transsulphuration enzymes is gained by the comparison to cystathionine beta-lyase from E. coli, indicating the mechanistic importance of a second substrate binding site for L-cysteine which leads to different chemical reaction types. 相似文献
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