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1.
The effect of anaerobiosis (N2 bubbling of the medium) or 10(-4) M dinitrophenol on the penetration of 0.5 mM Phe in snail and rat everted intestine, in 2 min and 30 min incubation periods, has been studied. The aerobic energy deficit inhibits the amino acid net entry in both species, but whereas the active transport is annulled in rat, snail intestine is capable of continuing to accumulate Phe against a gradient. The prolonged action (30 min of preincubation) of 1 mM ouabain inhibits 0.1 mM Phe and 0.1 mM galactose entry in snail intestine. Amino acid uptake is far higher than the one obtained in the absence of Na+, in which condition Phe keeps accumulating against a gradient in the tissue water. Galactose active transport, instead, becomes null in the presence of the glucoside or in the absence of Na+. One mM harmaline is able to inhibit the initial entry of galactose into the tissue, while higher than 5 mM concentrations are required to inhibit that of Phe. Results confirm that snail intestine is capable of easily carrying out active transport processes with energy from anaerobic origin. On the other hand Phe transport is less sensitive to the absence of Na+, presence of ouabain or harmaline than that of galactose, so that contrary to what has been observed for the sugar, the active transport of the amino acid is not annulled in any of the three conditions. 相似文献
2.
The effect of various coumarins on the active transport of galactose by small intestine in chick and rat was studied, using the in vivo technique of sucessive absorptions. A 10(-4) M concentration of the different coumarins inhibits the absorption of galactose in the chick. This effect persists in successive absorptions without coumarin. In rat, inhibition of galactose active transport by coumarins was observed at 10(-3) M concentration. 相似文献
3.
Sugar transport by sacs of everted intestine of snail have been measured in vitro at 30 degrees C. D-galactose, D-glucose and 3-O-methylglucose were actively transported against a concentration gradient from the mucosal to the serosal compartment. The transport of these sugars was inhibited by 5 times 10(-8) to 10(-6) M phlorizin. L-arabinose was also accumulated in the serosal compartment against a concentration gradient; in this case, transport was not affected by phlorizin. The snail intestine did not show any ability for D-fructose active transport but there was a clear uptake of this sugar by the tissue. The O2 uptake of the snail intestine was not significantly affected by the presence of either sugars or phlorizin. 相似文献
4.
1. It has been shown that most of the intracellular potassium of turtle heart can be replaced by cesium. 2. The uptake of cesium is reduced by the presence of either DNP 5.10(-5) M or ouabain 10(-6) M in the external medium. The presence of 10(-5) M ouabain markedly inhibits the uptake of cesium. 3. These results lead to the conclusion that the intracellular accumulation of cesium occurs via an active inward transport system wich operates with the simultaneous efflux of sodium. 相似文献
5.
Active transport of sugars (D-galactose, D-glucose, 3-0-methylglucose and L-arabinose) by sacs of everted intestine of snail (Cryptomphalus hortensis) was strongly inhibited, but not abolished, when all Na from the bathing solutions was substituted by K, Tris, Mg or Ca. Absence of Na produced also a marked inhibition of O2 consumption by the tissue. Omission of other cations (K, Ca, Mg), substituted by Tris, did not affect sugar transport or O2 uptake. Sodium seems to play a specific and important but not indispensable r?le in sugar active transport by snail intestine. Since anaerobiosis did not affect sugar transport, this Na role is independent of its effect on O2 uptake. 相似文献
6.
The net entry of galactose into the tissue of snail everted intestinal rings with 2 or 15 minute long incubation periods has been measured. With 10(-4) M phlorizin, the mediated transport is completely blocked while only the passive entry of sugar is produced. Lower concentrations of the glycoside partially inhibit transport according to competitive inhibition kinetics (K1 = 10(-7) M). The transport of galactose is Na+ dependent. In the absence of Na+, transport ceases and the sugar entry can be explained through simple diffusion. With 15 mM Na+ (control 71,4 mM) transport diminishes and a marked increase in the apparent Km with no changes in the Vmax is observed. One mM harmaline completely blocks galactose (0.5 mM) transport. One mM ouabain also makes transport null, but only after tissue preincubation with the inhibitor on the serosal side. 相似文献
7.
Patel N Khayat ZA Ruderman NB Klip A 《Biochemical and biophysical research communications》2001,285(4):1066-1070
2,4-dinitrophenol (DNP) compromises ATP production within the cell by disrupting the mitochondrial electron transport chain. The resulting loss of ATP leads to an increase in glucose uptake for anaerobic generation of ATP. In L6 skeletal muscle cells, DNP increases the rate of glucose uptake by twofold. We previously showed that DNP increases cell surface levels of glucose transporter 4 (GLUT4) and hexose uptake via a Ca2+-sensitive and conventional protein kinase C (cPKC)-dependent mechanism. Recently, 5' AMP-activated protein kinase (AMPK) has been proposed to mediate the stimulation of glucose uptake by energy stressors such as exercise and hypoxia. Changes in Ca2+ and cPKC have also been invoked in the stimulation of glucose uptake by exercise and hypoxia. Here we examine whether changes in cytosolic Ca2+ or cPKC lead to activation of AMPK. We show that treatment of L6 cells with DNP (0.5 mM) or hyperosmolar stress (mannitol, 0.6 M) increased AMPK activity by 3.5-fold. AMPK activation peaked by 10-15 min prior to maximal stimulation of glucose uptake. Intracellular Ca2+ chelation and cPKC inhibition prior to treatment with DNP and hyperosmolarity significantly reduced cell surface GLUT4 levels and hexose uptake but had no effect on AMPK activation. These results illustrate a break in the relationship between AMPK activation and glucose uptake in skeletal muscle cells. Activation of AMPK does not suffice to stimulate glucose uptake in response to DNP and hyperosmolarity. 相似文献
8.
Separation of phenylalanine transport events by using selective inhibitors, and identification of a specific uncoupler activity in Yersinia pestis.
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Phenylalanine transport in Yersinia pestis TJW was differentially inhibited by sulfhydryl blocking reagents, uncoupling agents, and respiratory inhibitors. Kinetic studies with potassium cyanide and sodium azide showed that these compounds have no immediate effect on the initial rate of phenylalanine transport, but have an immediate and severe inhibitory effect on the rate of oxygen uptake. Identical studies with p-chloromercuribenzoate (pCMB) and 2,4-dinitrophenol (DNP) showed that these compounds have an instantaneous and total inhibitory effect on phenylalanine transport. DNP stimulated oxygen uptake, and pCMB caused only a sluggish inhibiton of oxygen uptake. pCMB acted as a competitive inhibitor of phenylalanine transport, whereas DNP inhibitied noncompetitively. Arrenius plots of the initial rate of phenylalanine transport in pCMB- and DNP-treated cells showed that DNP alters the transition temperature of the phenylalanine transport system from 17 C for control cells to 12 C. DNP did not inhibit transport when cells were treated at temperatures of 2 to 10 C. PCMB did not alter the normal transition temperature and inhibited phenylalanine transport over a 2 to 30 C temperature range. Efflux induced by both pCMB and DNP were blocked by placing cells at low temperatures (2 to 20 C). Inhibition of adenosine 5'-triphosphate synthesis by DNP did not show any temperature sensitivity as did phenylalanine transport. These data indicate that: (i) respiration is not obligatory for active transport of phenylalanine in Y. pestis TJW; and (ii) pCMB inhibits transport activity by reacting with the sulfhydryl group(s) at the carrier binding site. The data show that the uncoupler, DNP, selectively alters a temperature-dependent property of phenylalanine transport, that is not related to uncoupling activity of DNP , and probably involves membrane lipid alterations. 相似文献
9.
The effect of phenformin on the absorption of D-glucose and D-galactose by hamster and rat intestine, was studied. Phenformin did not affect D-glucose absorption by rat intestine, but it inhibited at 10(-3) to 10(-2) M the absorption of D-glucose and D-galactose by hamster intestine. The inhibition was higher when D-glucose was tested. Phenformin also inhibited active accumulation of these sugars by rings of hamster small intestine, in vitro; this effect was greater when D-glucose was utilized. The drug inhibits the oxygen uptake in the tissue in the absence or in the presence of added substrate. Phenformin, as previously suggested, does not seem to act as a specific inhibitor on D-glucose transport, but most likely by its inhibitory effect on mitochondrial respiration. 相似文献
10.
The effect of p-chloromercuribenzoic acid (pCMB), either alone or in the presence of 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), on the 1 mM galactose absorption by in vivo perfused rat intestine has been studied. At 0.25 mM concentration, pCMB inhibits galactose absorption in about 32% but it does not modify the absorption of this sugar when the transport is blocked by 0.5 mM phlorizin, or that of the non-transportable monosaccharide derivative 2-deoxy-D-glucose. This shows that only the active transport component of galactose absorption is inhibited. A 2 min preexposure period is required for the inhibition to appear. The inhibition was not reversed by washing with saline solution even when it contained 0.5 mM dithioerythritol, 10 mM cysteine or 5 and 10 mM EDTA. The simultaneous exposure to 0.25 pCMB and 0.25 mM DTNB inhibits the total galactose entry in about 50%, an effect higher than the one exerted by each reagent separately and close to the one obtained with 0.5 mM phlorizin. Our results, in vivo, confirm the importance of the thiol groups in the cotransport of Na+ and sugar. As DTNB is an SH-reagent of lesser liposolubility than pCMB, the existence of two populations of sulfhydryl groups related to sugar transport which differ in their location within the brushborder membrane and in accessibility from the intestinal lumen, is suggested. 相似文献
11.
Tamás König András Kapus Balázs Sarkadi 《Journal of bioenergetics and biomembranes》1993,25(5):537-545
The effect of equisetin, an antibiotic produced byFusarium equiseti, has been studied on mitochondrial functions (respiration, ATPase, ion transport). Equisetin inhibits the DNP-stimulated ATPase activity of rat liver mitochondria and mitoplasts in a concentration-dependent manner; 50% inhibition is caused by about 8 nmol equisetin/mg protein. The antibiotic is without effect either on the ATPase activity of submitochondrial particles or on the purified F1-ATPase. It inhibits both the ADP- or DNP-activated oxygen uptake by mitochondria in the presence of glutamate + malate or succinate as substrates, but only the ADP-stimulated respiration is inhibited if the electron donors are TMPD + ascorbate. It does not affect the NADH or succinate oxidation of submitochondrial particles. Equisetin inhibits in a concentration-dependent manner the active Ca2+-uptake of mitochondria energized both by ATP or succinate without affecting the Ca2+-uniporter itself. The antibiotic inhibits the ATP-uptake by mitochondria (50% inhibition at about 8 nmol equisetin/mg protein) and the Pi and dicarboxylate carrier. It does not lower the membrane potential at least up to 200 nmol/mg protein concentration. The data presented in this paper indicate that equisetin specifically inhibits the substrate anion carriers of the mitochondrial inner membrane.Abbreviations EGTA
ethyleneglycol bis/-aminoethylether/-N, N-tetraacetic acid
- DNP
2, 4-dinitrophenol
- TMPD
N,N,N,N,tetramethyl-p-phenylenediamine
- CCP
carbonylcyanide-m-chlorophenyl hydrazone
- TPP
tetraphenyl-phosphonium
- Hepes
/4,(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid/ 相似文献
12.
Ionomycin, a recently discovered calcium ionophore, inhibits the ATP-dependent active Ca2+ transport of rabbit sarcoplasmic reticulum vesicles at concentrations as low as 10(-8) to 10(-6) M. The effect is due to an increase in the Ca2+ permeability of the membrane which is also observed on liposomes. The inhibition of Ca2+ uptake is accompanied by an increase in the Ca2+-sensitive ATPase activity of sarcoplasmic reticulum vesicles. 相似文献
13.
Involvement of PKC and PKA in the inhibitory effect of leptin on intestinal galactose absorption 总被引:1,自引:0,他引:1
Barrenetxe J Sainz N Barber A Lostao MP 《Biochemical and biophysical research communications》2004,317(3):717-721
Studies from our laboratory have demonstrated that leptin inhibits galactose absorption in vitro by acting on the Na(+)/glucose cotransporter SGLT1. Since PKC and PKA are involved in the regulation of SGLT1 and leptin is able to activate these kinases, we have investigated the possible implication of PKC and PKA in the inhibition of sugar absorption by leptin in rat small intestinal rings. Inhibition of 1 mM galactose uptake by 0.2 nM leptin is blocked by 2 microM chelerythrine, a PKC inhibitor, which by itself does not affect galactose uptake. However, 1 microM H-89, a PKA inhibitor, inhibits galactose uptake and does not block leptin inhibition. Biochemical assays show that the inhibitory effect of leptin is accompanied by a approximately 2-fold increase in PKA and PKC activity. These findings indicate that the activation of PKC is more relevant than PKA activation in the inhibition of galactose absorption by leptin. 相似文献
14.
Hypoxia-induced inhibition of whole cell membrane currents and ion transport of A549 cells 总被引:1,自引:0,他引:1
Karle C Gehrig T Wodopia R Höschele S Kreye VA Katus HA Bärtsch P Mairbäurl H 《American journal of physiology. Lung cellular and molecular physiology》2004,286(6):L1154-L1160
In excitable cells, hypoxia inhibits K channels, causes membrane depolarization, and initiates complex adaptive mechanisms. It is unclear whether K channels of alveolar epithelial cells reveal a similar response to hypoxia. A549 cells were exposed to hypoxia during whole cell patch-clamp measurements. Hypoxia reversibly inhibited a voltage-dependent outward current, consistent with a K current, because tetraethylamonium (TEA; 10 mM) abolished this effect; however, iberiotoxin (0.1 microM) does not. In normoxia, TEA and iberiotoxin inhibited whole cell current (-35%), whereas the K-channel inhibitors glibenclamide (1 microM), barium (1 mM), chromanol B293 (10 microM), and 4-aminopyridine (1 mM) were ineffective. (86)Rb uptake was measured to see whether K-channel modulation also affected transport activity. TEA, iberiotoxin, and 4-h hypoxia (1.5% O(2)) inhibited total (86)Rb uptake by 40, 20, and 35%, respectively. Increased extracellular K also inhibited (86)Rb uptake in a dose-dependent way. The K-channel opener 1-ethyl-2-benzimidazolinone (1 mM) increased (86)Rb uptake by 120% in normoxic and hypoxic cells by activation of Na-K pumps (+60%) and Na-K-2Cl cotransport (+170%). However, hypoxic transport inhibition was also seen in the presence of 1-ethyl-2-benzimidazolinone, TEA, and iberiotoxin. These results indicate that hypoxia, membrane depolarization, and K-channel inhibition decrease whole cell membrane currents and transport activity. It appears, therefore, that a hypoxia-induced change in membrane conductance and membrane potential might be a link between hypoxia and alveolar ion transport inhibition. 相似文献
15.
The dose-dependent effect of hydrogen peroxide on snail neuromembrane chemosensitivity was studied by means of standard voltage-clamp method. Short-term exposure (7 min) of neurons to H(2)O(2) (10(-11)-10(-4) M) caused dose-dependent depression of Acetylcholine (Ach)-induced ionic currents in the membrane. The H(2)O(2)-induced depression of Ach-sensitivity of membrane was more pronounced in K(+)-free solution than in normal physiological solution and it disappeared in cold medium (5 degrees C). The H(2)O(2) (10(-11)-10(-4) M) decreased membrane electrical conductivity and cell volume. The dose-dependent decrease in Ach-sensitivity of the snail neuromembrane by H(2)O(2) may be due to a decrease in the number of functionally active membrane receptors caused by a decrease in membrane active surface. H(2)O(2)-induced decrease in Ach-sensitivity has a metabolic but Na(+)-K(+) pump independent character, the nature of which is the subject for current investigation. 相似文献
16.
We report the kinetic characteristics for D-galactose, 2-deoxy-D-glucose and 3-O-methyl-D-glucose transport in a galactokinase null-allele mutant of a Chinese hamster V79 cell line. GalKl cells exhibited a Km and Vmax for D-galactose, 2-deoxy-D-glucose, and 3-O-methyl-D-glucose transport of 8.6 +/- 2.6 mM and 26.1 +/- 7.2 nmol/mg p/min, 4.1 +/- 1.2 mM and 40.3 +/- 9.5 nmol/mg p/min, and 7.01 +/- .85 mM and 11.6 +/- 4.8 nmol/mg p/30 s, respectively. Nonsaturable hexose uptake was determined using cytochalasin B inhibition of galactose uptake (89.6 +/- 3.7% of galactose uptake was cytochalasin B inhibitable) and L-glucose uptake (7.5% of the galactose uptake). D-Galactose was not metabolized and effluxed rapidly from preloaded cells. The Kls for the inhibition of D-galactose transport were 4.5 +/- 2.5 mM for D-glucose, 7.0 +/- 2.0 mM for 2-deoxy-D-glucose, 6 mM for 2-deoxy-D-galactose and 6.0 +/- 0.6 mM for 3-O-methyl-D-glucose. This indicates the operation of a single common carrier. The hexose transport rate decreased 50-60% after 24 h serum deprivation. Addition of insulin was shown to increase hexose transport (more than twofold) in serum-deprived cells. Hexose transport rates increased substantially in glucose-deprived, D-fructose- or D-galactose-fed cells as compared to glucose-fed cells. Since GalKl does not metabolize galactose, the hexose transport increases induced by feeding cells galactose suggest that carrier interaction with ligand is not a significant factor in transport regulation in GalKl. The kinetic and regulatory characteristics of D-galactose transport in the GalKl cell line indicate that this system is a good model to study sugar transport from a mechanistic and regulatory point of view. 相似文献
17.
Erik Dybing 《生物化学与生物物理学报:生物膜》1974,373(1):100-105
Cycloheximide preincubation inhibits hypoxanthine uptake into the acid-soluble fractions of cultured rat hepatoma cells (MH1C1) and human skin epithelial cells (NCTC 2544, HE cells) in a time- and dose-dependent manner 50% inhibition is seen after 4 h preincubation with 10?4 M cycloheximide of MH1C1 cells and after 2.5 h of HE cells. Adenine uptake is much less affected, after 10 h preincubation with 10?4 M cycloheximide it was reduced to 83% and 67% of controls in MH1C1 cells and HE cells respectively. Cycloheximide inhibits hypoxanthine uptake in a dose-dependent manner above 10?7 M, with 50% inhibition in MH1C1 cells at 4 · 10?7 M after 12 h preincubation and at 10-6 M in HE cells after 6 h preincubation. Puromycin mimics the action of cycloheximide. The inhibition of hypoxanthine uptke is not caused by reduction of the activity of hypoxanthine phosphoribosyltransferase in the two cell lines. 10?4 M cycloheximide preincubation for 10 h does not significantly reduce the uptake of the two non-metabolizable amino acids α-aminoisobutyric acid or 1-aminocyclopentane-1-carboxylic acid (cycloleucine). It is suggested that cycloheximide inhibits the synthesis of a rapidly turning over the protein involved in hypoxanthine transport. 相似文献
18.
Role of microtubules in insulin and glucagon stimulation of amino acid transport in isolated rat hepatocytes 总被引:4,自引:0,他引:4
The effects of the microtubule inhibitor, colchicine, on insulin or glucagon stimulation of alpha-amino[1-14C]-isobutyric acid (AIB) transport were investigated in isolated hepatocytes from normal fed rats. Under all conditions tested, AIB uptake appeared to occur through two components of transport: a low affinity (Km approximately 50 mM) component and a high affinity (Km approximately 1 mM) component. Within 2 h of incubation, insulin and glucagon, at maximal concentrations, increase AIB (0.1 mM) uptake by 2- to 3-fold and 4- to 6-fold, respectively. Colchicine, at the low concentration of 5 X 10(-7) M, slightly reduces basal AIB transport, decreases by 80% the simulatory effect of insulin, and diminishes by 40% the stimulatory effect of either glucagon or dibutyryl cAMP. Kinetic analysis of AIB influx indicates that the drug inhibits the increase in Vmax of a high affinity (Km approximately 1 mM) component of transport stimulated by insulin or glucagon, without affecting the kinetic parameters of a low affinity component of transport (Km approximately 50 mM). Various short term hormonal effects of insulin and glucagon (changes in glucose, urea, and lactate production) were found not to be modified by the drug. Vinblastine elicits similar changes as colchicine on AIB uptake. Lumicolchicine, a colchicine analogue that does not bind to tubulin, has no effect. The concentration of colchicine (10(-7) M) required for half-maximal inhibition of hormone-stimulated AIB transport is in the appropriate range for specific microtubule disruption. These data suggest that microtubules are involved in the regulation of the insulin or glucagon stimulation of AIB transport in isolated rat hepatocytes. 相似文献
19.
J Nargeot 《Journal de physiologie》1976,72(2):171-180
Current clamp studies showed that after 10 minutes under DNP 10(-4) M the membrane potential does not change significantly while an important shortening of the action potential duration and a diminished amplitude are observed. Voltage clamp studies have been performed on the slow inward and delayed outward currents. DNP 10(-4) M induced a marked decrease of the slow inward current related to the reduction in both conductance and driving force, and a decrease in the amplitude of the delayed current. The decrease of the slow inward current seems to be mainly responsible for the suppression of the plateau of the action potential during metabolic inhibition. 相似文献
20.
In coupled mitochondria of maize, inorganic pyrophosphate has no effect on electron transport whereas it competitively inhibits state 3 (with addition of ADP) respiration. The degree of inhibition depends on the ADP concentration in the reaction medium. At 150 and 30O μM ADP, the inhibition constant (Ki) has a value of 1.1 × 10−4 M. Pyrophosphate either does not penetrate throucvh the membranes or penetrates through them in only very small amounts. It does not inhibit the exchange 32PiPi; however, it undergoes an exchange with ADP (2 nmol PPi/mg protein for 10 min at 30°). 相似文献