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1.
《生物化学与生物物理学报:生物膜》1980,598(1):127-133
2,4-Fluorodinitrobenzene and 2,3-butanedione, which irreversibly inactivate the glucose transfer system of erythrocytes, have been used as probes to determine whether the substrate site and inner and outer sites for reversible inhibitors are located in the same or different regions of the carrier. Inhibitors bound at an inhibition site exposed in the inward-facing but not the outwardfacing form of the carrier (cytochalasin B, androstendione and androstandione) protect the transport system against inactivation by 2,4-fluorodinitrobenzene. Inhibitors bound at an external inhibition site (phloretin) and substrates bound at the transfer site do not protect. In contrast inactivation by 2,3-butanedione is slightly accelerated by internally bound inhibitors, while substrates and substrate analogs bound at the transfer site protect the system. It is shown that fluorodinitrobenzene reacts in the inner inhibition site and butanedione in the substrate site; and further that these sites may be separate binding areas in the carrier linked by allosteric interaction. The consequence of this linkage is that binding of a ligand at the substrate site precludes binding of another ligand at the internal or external inhibition site. 相似文献
2.
There are several reports on the oxidation of external NADH by an exogenous NADH dehydrogenase in the outer leaflet of the inner membrane of rat heart mitochondria. Until now, however, little was known about its physiological role in cellular metabolism. The present work shows that carvedilol (?1-[carbazolyl-(4)-oxy]-3-[2-methoxyphenoxyethyl)amino]-pro - panol-(2)?) is a specific inhibitor of an exogenous NADH dehydrogenase in rat heart mitochondria. Carvedilol does not affect oxygen consumption linked to the oxidation of succinate and internal NADH. It is also demonstrated that the inhibition of exogenous NADH dehydrogenase by carvedilol is accompanied by the inhibition of alkalinization of the external medium. In contrast to the addition of glutamate/malate or succinate, exogenous NADH does not generate a membrane potential in rat heart mitochondria, as observed with a TPP(+) electrode. It is also demonstrated that the oxygen consumption linked to NADH oxidation is not due to permeabilized mitochondria, but to actual oxidase activity in the inner membrane. The enzyme has a K(m) for NADH of 13 microM. Carvedilol is a noncompetitive inhibitor of this external NADH dehydrogenase with a K(i) of 15 microM. Carvedilol is the first inhibitor described to this organospecific enzyme. Since this enzyme was demonstrated to play a key role in the cardiotoxicity of anticancer drugs of the anthracycline family (e.g., adriamycin), we may suggest that the administration of carvedilol to tumor patients treated with adriamycin might be of great help in the prevention of the cardioselective toxicity of this antibiotic. 相似文献
3.
The ROMK (Kir1.1) family of epithelial K channels can be inactivated by a combination of low internal pH and low external K, such that alkalization does not reopen the channels unless external K is elevated. Previous work suggested that this inactivation results from an allosteric interaction between an inner pH gate and an outer K sensor, and could be described by a simple three-state kinetic model. In the present study, we report that a sustained depolarization slowly inactivated (half-time = 10-15 min) ROMK channels that had been engineered for increased affinity to internal polyamines. Furthermore, this inactivation occurred at external [K] < or =1 mM in ROMK mutants whose inner pH gate was constitutively open (ROMK2-K61M mutation). Both pH and voltage inactivation depended on external K in a manner reminiscent of C-type inactivation, but having a much slower time course. Replacement of ROMK extracellular loop residues by Kir2.1 homologous residues attenuated or abolished this inactivation. These results are consistent with the hypothesis that there are (at least) two separate closure processes in these channels: an inner pH-regulated gate, and an outer (inactivation) gate, where the latter is modulated by both voltage and external [K]. 相似文献
4.
《Journal of Russian & East European Psychology》2013,51(4):46-53
The time has come to proceed from forms of givenness of the word to forms of the word as such. They can, if you like, be called external and inner structures. Humboldt, however, preferred to speak of the external and inner forms of the language. Shpet adopted precisely this distinction. Why did this problem interest Shpet? Already in [Appearance and sense], he had set the task of returning to the source of pretheoretical, living science. Shpet wrote that the outer cover of words and logical expressions obscure the objectified meaning and that it was necessary to remove another cover from the objectified sign so as to grasp a certain genuine intimacy, and in it the fullness of being (Shpet, 1914. Pp. 5-6). We shall keep in mind this major undertaking posed by this scientist. The existence of the inner form of words should not come as a surprise. That same year (1914) Ortega y Gasset wrote that material objects have a third dimension. However, we cannot see or touch it: "For just as depth needs a surface beneath which to be concealed, the surface, or outer cover, in order to be so, needs something over which to spread, covering it" (Ortega y Gasset, 2000. Pp. 62-63). 相似文献
5.
Protein import into mitochondria: the requirement for external ATP is precursor-specific whereas intramitochondrial ATP is universally needed for translocation into the matrix. 总被引:17,自引:4,他引:13
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ATP is needed for the import of precursor proteins into mitochondria. However, the role of ATP and its site of action have been unclear. We have now investigated the ATP requirements for protein import into the mitochondrial matrix. These experiments employed an in vitro system that allowed ATP levels to be manipulated both inside and outside the mitochondrial inner membrane. Our results indicate that there are two distinct ATP requirements for mitochondrial protein import. ATP in the matrix is always needed for complete import of precursor proteins into this compartment, even when the precursors are presented to mitochondria in an unfolded conformation. In contrast, the requirement for external ATP is precursor-specific; depletion of external ATP strongly inhibits import of some precursors but has little or no effect with other precursors. A requirement for external ATP can often be overcome by denaturing the precursor with urea. We suggest that external ATP promotes the release of precursors from cytosolic chaperones, whereas matrix ATP drives protein translocation across the inner membrane. 相似文献
6.
Vilbig RL Sarkar A Zischkau J Knepper MA Pisitkun T 《American journal of physiology. Renal physiology》2012,303(3):F366-F372
Concentrating models of the renal inner medulla can be classified according to external free-energy balance into passive models (positive values) and models that require an external energy source (negative values). Here we introduce an online computational tool that implements the equations of Stephenson and colleagues (Stephenson JL, Tewarson RP, Mejia R. Proc Natl Acad Sci USA 71: 1618-1622, 1974) to calculate external free-energy balance at steady state for the inner medulla (http://helixweb.nih.gov/ESBL/FreeEnergy). Here "external free-energy balance" means the sum of free-energy flows in all streams entering and leaving the inner medulla. The program first assures steady-state mass balance for all components and then tallies net external free-energy balance for the selected flow conditions. Its use is illustrated by calculating external free-energy balance for an example of the passive concentrating model taken from the original paper by Kokko and Rector (Kokko JP, Rector FC Jr. Kidney Int 2: 214-223, 1972). 相似文献
7.
Lemeshko VV 《Archives of biochemistry and biophysics》2001,388(1):60-66
The possibility of direct oxidation of external NADH in rat liver mitochondria and of the inner membrane potential generation in this process is still not clear. In the present work, the energy-dependent swelling of mitochondria in the medium containing valinomycin and potassium acetate was measured as one of the main criteria of the proton-motive force generation by complex III, complex IV, and both complexes III and IV of the respiratory chain. Mitochondria swelling induced by external NADH oxidation was compared with that induced by succinate or ferrocyanide oxidation, or by electron transport from succinate to ferricyanide. Mitochondria swelling, nearly equal to that promoted by ferrocyanide oxidation, was observed under external NADH oxidation, but only after the outer mitochondrial membrane was ruptured as a result of the swelling-contraction cycle, caused by succinate oxidation and its subsequent inhibition. In this case, significantly accelerated intermembrane electron transport and well-detected inner membrane potential generation, in addition to mitochondria swelling, were also observed. Presented results suggest that exogenous NADH and cytochrome c do not support the inner membrane potential generation in intact rat liver mitochondria, because the external NADH-cytochrome c reductase system, oriented in the outer mitochondrial membrane toward the cytoplasm, is inaccessible for endogenous cytochrome c reduction; as well, the inner membrane cytochrome c oxidase is inaccessible for exogenous cytochrome c oxidation. 相似文献
8.
Cytochrome o type oxidase from Escherichia coli. Characterization of the enzyme and mechanism of electrochemical proton gradient generation 总被引:8,自引:0,他引:8
Cytochrome o type oxidase purified from the membrane of Escherichia coli consists of four polypeptides (Mr 66000, 35000, 22000, and 17000), and the monomeric form predominates in octyl beta-D-glucopyranoside. The oxidase complex contains two b-type cytochromes (b-558 and b-563) and 2 mol of heme/mol of enzyme. Cytochrome o utilizes ubiquinol-1 and a number of other artificial electron donors as substrates but does not oxidize reduced cytochrome c or ferrocyanide. Activity is highly dependent upon exogenous phospholipids and/or Tween 20, and the quinone analogues 2-heptyl-4-hydroxyquinoline N-oxide and 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole are potent inhibitors. Proteoliposomes were formed by detergent dilution or dialysis in the presence of the oxidase and phospholipids, followed by freeze-thaw/sonication. Vesicles formed by this means are unilamellar and contain a random distribution of 85-90-A intramembranous particles on the convex and concave fracture surfaces. During oxidase turnover, the reconstituted system generates a proton electrochemical gradient (interior negative and alkaline) of -115 to -140 mV; however, respiratory control is minimal (i.e., respiratory control ratios of about 1.5 are observed). By using a glass electrode to measure changes in external pH and the fluorescence of entrapped 8-hydroxy-1,3,6-pyrenetrisulfonate to measure changes in internal pH, it is apparent that during ubiquinol oxidation, protons are released on the external surface of the membrane and consumed on the internal surface. In contrast, with N,N,N',-N'-tetramethyl-p-phenylenediamine, an electron donor that carries few protons at neutral pH, little change in external pH is observed until the protonophore carbonyl cyanide m-chlorophenylhydrazone is added, at which point the medium becomes alkaline. The results taken as a whole are consistent with the concept that oxidase turnover generates an electrical potential (interior negative) due to vectorial electron flow from the outer to the inner surface of the membrane. The pH gradient (interior alkaline), on the other hand, appears to result from scalar (i.e., nonvectorial) reactions that consume and release protons at the inner and/or outer surfaces of the membrane, respectively. In other words, cytochrome o oxidase from Escherichia coli does not appear to catalyze vectorial proton translocation. 相似文献
9.
Entire hydras or tentacles were prepared for electron microscopy as described in the preceding paper. The stenotele capsule has been observed to be composed of an external membrane, a thick chitinous or keratin layer, and an inner membrane. A sac-like extension of the capsular wall into the capsule bears spines and stylets on its inner surface and evagination of this structure occurs on discharge. Profiles of tubular or membranous structures often are seen within the capsules of resting stenoteles. These structures are presumably related to the external filament. The spines often reveal a flattened aspect which suggests that at least some of them might more accurately be called "vanes." A cnidocil has been found to accompany each stenotele. This study revealed several aspects of the developmental stages of stenoteles: A vacuole is formed which is nearly surrounded by the nematocyte nucleus. The vacuole content changes in density and a capsular wall is formed at the periphery of the vacuole. Tubules differentiate from the capsular matrix, and spines and stylets develop somewhat later. An operculum is formed from the nematocyte cytoplasm. 相似文献
10.
Entire hydras or tentacles were prepared for electron microscopy as described in the preceding paper. The stenotele capsule has been observed to be composed of an external membrane, a thick chitinous or keratin layer, and an inner membrane. A sac-like extension of the capsular wall into the capsule bears spines and stylets on its inner surface and evagination of this structure occurs on discharge. Profiles of tubular or membranous structures often are seen within the capsules of resting stenoteles. These structures are presumably related to the external filament. The spines often reveal a flattened aspect which suggests that at least some of them might more accurately be called "vanes." A cnidocil has been found to accompany each stenotele. This study revealed several aspects of the developmental stages of stenoteles: A vacuole is formed which is nearly surrounded by the nematocyte nucleus. The vacuole content changes in density and a capsular wall is formed at the periphery of the vacuole. Tubules differentiate from the capsular matrix, and spines and stylets develop somewhat later. An operculum is formed from the nematocyte cytoplasm. 相似文献
11.
A case of atresia of the external acoustic meatus is presented from a prehistoric Late Woodland site in Iowa. The affected individual was a female aged 30 to 40 years. The left temporal bone displayed a thin bony plate occluding the external acoustic canal. No evidence of middle ear or inner ear malformation was observed on the axial tomographs, although the external canal was shortened. The individual is diagnosed as having a mild case of congenital atresia. The report extends the geographical and temporal distributions of this archaeologically rare condition. 相似文献
12.
C. Andersen 《Reviews of Physiology, Biochemistry and Pharmacology》2003,147(1):122-165
For translocation across the cell envelope of Gram-negative bacteria, substances have to overcome two permeability barriers,
the inner and outer membrane. Channel-tunnels are outer membrane proteins, which are central to two distinct export systems:
the type I secretion system exporting proteins such as toxins or proteases, and efflux pumps discharging antibiotics, dyes,
or heavy metals and thus mediating drug resistance. Protein secretion is driven by an inner membrane ATP-binding cassette
(ABC) transporter while drug efflux occurs via an inner membrane proton antiporter. Both inner membrane transporters are associated
with a periplasmic accessory protein that recruits an outer membrane channel-tunnel to form a functional export complex. Prototypes
of these export systems are the hemolysin secretion system and the AcrAB/TolC drug efflux pump of Escherichia coli, which both employ TolC as an outer membrane component. Its remarkable conduit-like structure, protruding 100 ? into the
periplasmic space, reveals how both systems are capable of transporting substrates across both membranes directly from the
cytosol into the external environment. Proteins of the channel-tunnel family are widespread within Gram-negative bacteria.
Their involvement in drug resistance and in secretion of pathogenic factors makes them an interesting system for further studies.
Understanding the mechanism of the different export apparatus could help to develop new drugs, which block the efflux pumps
or the secretion system.
Electronic Publication 相似文献
13.
Oocysts and sporocysts of Eimeria funduli sp. n. are described from the Gulf killifish, Fundulus grandis, on the basis of light microscopy, transmission and scanning electron miscroscopy, and location in the liver of infected hosts. The spherical sporulated oocysts of E. funduli isolated from liver tissue measure 20-31 (25) micrometer across with ovoid sporocysts 9-11 X 5-7 (10 X 6) micrometer. A micropyle, polar granule, and oocyst residuum are absent, but sporocysts have Stieda and substieda bodies, a few residual granules, and 10-25 (15) unique projecting structures with expanded distal portions that we term "sporopodia". Sporopodia 1-3 (2) micrometer high support a transparent membrane that completely surrounds the sporocyst. Sporozoites have one large posterior refractile body. Ultrastructurally, the oocyst wall consists of two thin layers of granular material: an electron-dense outer layer with a rough external surface and an electron-lucent inner one of approximately equal thickness. One or two unit membranes line the inner surface of the inner layer. Each layer is 40-60 (55) nm thick. The sporocyst wall, 78-130 (110) nm thick, consists of an electron-lucent material with the outer surface being more electron dense and giving rise to osmiophilic sporopodia; closely associated with these and the outer surface are one or two unit membranes. A thin osmiophilic layer of fine granular material lines the inner surface. 相似文献
14.
Chigri F Soll J Vothknecht UC 《The Plant journal : for cell and molecular biology》2005,42(6):821-831
The majority of chloroplast proteins is nuclear-encoded and therefore synthesized on cytosolic ribosomes. In order to enter the chloroplast, these proteins have to cross the double-membrane surrounding the organelle. This is achieved by means of two hetero-oligomeric protein complexes in the outer and inner envelope, the Toc and Tic translocon. The process of chloroplast import is highly regulated on both sides of the envelope membranes. Our studies indicate the existence of an undescribed mode of control for this process so far, at the same time providing further evidence that the chloroplast is integrated into the calcium-signalling network of the cell. In pea chloroplasts, the calmodulin inhibitor Ophiobolin A as well as the calcium ionophores A23187 and Ionomycin affect the translocation of those chloroplast proteins that are imported with an N-terminal cleavable presequence. Import of these proteins is inhibited in a concentration-dependent manner. Addition of external calmodulin or calcium can counter the effect of these inhibitors. Translocation of chloroplast proteins that do not possess a cleavable transit peptide, that is outer envelope proteins or the inner envelope protein Tic32, is not affected. These results suggest that the import of a certain subset of chloroplast proteins is regulated by calcium. Our studies furthermore indicate that this regulation occurs downstream of the Toc translocon either within the intermembrane space or at the inner envelope translocon. A potential promoter of the calcium regulation is calmodulin, a protein well known as part of the plant's calcium signalling system. 相似文献
15.
Using 86Rb+ as a marker for K+ permeability, we find that extracellular Ca-EGTA influences the rate of 86Rb+ efflux from erythrocyte ghosts preloaded with 86Rb+ and "buffered" Ca2+. At an internal free Ca2+, where the rate of 86Rb+ efflux is minimal and uninfluenced by either external EGTA or external Ca2+, external Ca-EGTA at 0.2-0.5 mM can raise the flux rate to as high as can be attained by raising internal Ca2+, in the presence of an excess externally either of Ca2+ or of EGTA. Higher concentrations of Ca-EGTA (up to 1-2 mM) diminish the flux rate. External Ca-EDTA or Mg-EDTA can substitute for Ca-EGTA in enhancing and suppressing flux rate. The peak rate is insensitive to external free Ca2+ but depends on internal Ca2+; internal Mg-EDTA does not substitute for internal Ca-EGTA. Thus, the erythrocyte membrane is asymmetric with respect to its interaction with Ca2+ and Ca-EGTA. Also, 22Na+ does not substitute for 86Rb+. The peak rate of 86Rb+ flux produced by external Ca-EGTA is diminished by chlorpromazine (0.1 mM) and augmented by 1-propranolol (25 microM), in the same way as the rate produced by increasing internal Ca2+. The results suggest that external Ca-EGTA enhances the affinity of internal Ca2+ for its receptor(s) which operate the K+-gate at the inner surface of the membrane. At external concentrations of Ca-EGTA above 1-2 mM, 86Rb+ flux rate again rises with increase of Ca-EGTA. This phenomenon does not depend upon internal Ca2+, is not affected by chlorpromazine or by 1-propranolol, and is associated with an enhanced permeability to 22Na+, inulin, and haemoglobin. 相似文献
16.
Mireille Saint-Macary Bernard Foucher 《Biochemical and biophysical research communications》1980,96(1):457-462
The binding of malonate to the external face of the mitochondrial inner membrane has been investigated by using mitoplasts and on the opposite face, by using inside-out oriented vesicles prepared from sonicated mitoplasts. The external face of the inner membrane displays a single class of binding sites whereas two classes are observed in vesicles. In trypsin treated vesicles, only high-affinity sites are evidenced. The disappearance of the low affinity sites is correlatively related with the loss of succinate dehydrogenase activity. Both high-affinity sites of mitoplasts and vesicles are sensitive to the presence of malate; they are also masked by 2-butylmalonate, phosphate, citrate and mercurials. Our results suggest that the internal and external high-affinity sites of the inner membrane are involved in the dicarboxylate transport system. 相似文献
17.
Påhlman IL Larsson C Averét N Bunoust O Boubekeur S Gustafsson L Rigoulet M 《The Journal of biological chemistry》2002,277(31):27991-27995
In the yeast Saccharomyces cerevisiae, the two most important systems for conveying excess cytosolic NADH to the mitochondrial respiratory chain are external NADH dehydrogenase (Nde1p/Nde2p) and the glycerol-3-phosphate dehydrogenase shuttle. In the latter system, NADH is oxidized to NAD+ and dihydroxyacetone phosphate is reduced to glycerol 3-phosphate by the cytosolic Gpd1p; glycerol 3-phosphate gives two electrons to the respiratory chain via mitochondrial glycerol-3-phosphate dehydrogenase (Gut2p)-regenerating dihydroxyacetone phosphate. Both Nde1p/Nde2p and Gut2p are located in the inner mitochondrial membrane with catalytic sites facing the intermembranal space. In this study, we showed kinetic interactions between these two enzymes. First, deletion of either one of the external dehydrogenases caused an increase in the efficiency of the remaining enzyme. Second, the activation of NADH dehydrogenase inhibited the Gut2p in such a manner that, at a saturating concentration of NADH, glycerol 3-phosphate is not used as respiratory substrate. This effect was not a consequence of a direct action of NADH on Gut2p activity because both NADH dehydrogenase and its substrate were needed for Gut2p inhibition. This kinetic regulation of the activity of an enzyme as a function of the rate of another having a similar physiological function may be allowed by their association into the same supramolecular complex in the inner membrane. The physiological consequences of this regulation are discussed. 相似文献
18.
在不利的环境条件下,枝角类中有一部分种类可以形成卵鞍(ephippium),内含休眠卵。本文应用扫描电镜和透射电镜对隆线溞的卵鞍进行了超微结构的研究。研究表明:卵鞍外面大部分略呈浅的蜂窝状,内面则排布着多数卵石状小突起。卵鞍分为内外两层,两层的超微结构截然不同;各层又可分为三小层。 相似文献
19.
Rachidi M Lopes C Gassanova S Sinet PM Vekemans M Attie T Delezoide AL Delabar JM 《Mechanisms of development》2000,93(1-2):189-193
The TPRD gene (tetratricopeptide (TPR) containing Down syndrome gene) is one of the candidate genes in the Down syndrome chromosomal region-1. Duplication of this gene may be the cause of major phenotypic features of Down syndrome. Here we show that the TPRD expression is developmentally regulated during human embryogenesis. At the earliest stages of development (Carnegie 8-12) TPRD expression is ubiquitous. At later developmental stages (Carnegie stages 14, 16 and 18), it becomes restricted to the nervous system, as is the case for the mtprd gene during mouse development. We extended our analysis of TPRD expression during fetal development of the human nervous system (13, 22 and 24 weeks). A new oblique illumination technique was used to compare signal intensity and cell density. Some regions of the nervous system such as the external cortical layers of the brain, and the inner neuroblastic layer of the eye, strongly express the TPRD gene. 相似文献
20.
Response of the Frog Skin to Steady-State Voltage Clamping : II. The active pathway 总被引:4,自引:4,他引:0
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Active Na transport across frog skin was separated from passive Na movement utilizing urea influx as a measure of passive (shunt) permeability. In this manner, the response of the overall active Na transport system to an applied potential was determined over a range from +200 mV to -100 mV. Active Na transport displays saturation as a function of applied potential, and both the level of saturation and the potential at which it is achieved are functions of the Na concentration in the external solution. The saturation with potential appears to involve a different step in the transport process than the saturation of Na flux as a function of external Na concentration. The observations can be qualitatively described by either a one-barrier or two-barrier model of the Na transport system. 相似文献