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Biomedical subjects

F L Crane

Publications and source records attributed to F L Crane.

At least 73 records · Page 4Linked to original sources
PubMed1987

Retinoic acid inhibition of transplasmalemma diferric transferrin reductase.

All trans retinoic acid inhibited diferric transferrin reduction by HeLa cells. The NADH diferric transferrin reductase activity of isolated liver plasma membranes was also inhibited by retinoic acid. Retinol and retinyl acetate had very little effect. Transplasma membrane ferricyanide reduction by HeLa cells and NADH ferricyanide reductase of liver plasma membrane was also inhibited by retinoic acid, therefore the inhibition was in the electron transport system and not at the transferrin receptor. Since the transmembrane electron transport has been shown to stimulate cell growth, the growth inhibition by retinoic acid thus may be based on inhibition of the NADH diferric transferrin reductase.

Animals↗
PubMed1987

Diferric transferrin reduction stimulates the Na+/H+ antiport of HeLa cells.

Proton release from HeLa cells is stimulated by external oxidants for the transplasmalemma electron transport enzymes. These oxidants, such as ferricyanide and diferric transferrin, also stimulate cell growth. We now present evidence that proton release associated with the reduction of ferricyanide and diferric transferrin is through the Na+/H+ antiport. The stoichiometry of H+/e- release with diferric transferrin is over 50 to 1, which is greater than expected for oxidation of a protonated transmembrane electron carrier. Diferric transferrin induced proton release depends on external sodium and is inhibited by amiloride. Proton release is also inhibited when diferric transferrin reduction is inhibited by apotransferrin. A tightly coupled association between the redox system and the antiport is shown by sodium dependence and amiloride inhibition of diferric transferrin reduction. The results indicate a new role for ferric transferrin in growth stimulation by activation of the sodium-proton antiport.

Carrier Proteins↗
PubMed1987

Ruthenium ammine complexes as electron acceptors for growth stimulation by plasma membrane electron transport.

Ammineruthenium(III) complexes have been found to act as electron acceptors for the transplasmalemma electron transport system of animal cells. The active complexes hexaammineruthenium(III), pyridine pentammineruthenium(III), and chloropentaammineruthenium(III) range in redox potential (E'0) from 305 to -42 mV. These compounds also act as electron acceptors for the NADH dehydrogenase of isolated plasma membranes. Stimulation of HeLa cell growth, in the absence of calf serum, by these compounds provides evidence that growth stimulation by the transplasma membrane electron transport system is not entirely based on reduction and uptake of iron.

Aminopyridines↗
PubMed1987

Involvement of transferrin in the reduction of iron by the transplasma membrane electron transport system.

Nonpermeable electron acceptors can be reduced by a transplasma membrane electron transport system in suspensions of intact cells. Here we report that diferric transferrin is reduced by HeLa S3 cells. The reduction is recorded spectrophotometrically as the formation of the ferrous complex of bathophenanthroline disulfonate. Ferric ammonium citrate can also be used as an electron acceptor and the presence of low concentrations of diferric transferrin greatly stimulates the reduction of trivalent iron under these conditions. Likewise very low concentrations of ferricyanide, which does not give rise to a ferrous bathophenanthroline disulfonate complex formation, have a strong stimulatory effect on the complex formation when ferric ammonium citrate is the source of ferric iron. Apotransferrin is a potent inhibitor of the reaction. The inhibition occurs at the concentration necessary for complete occupancy of the transferrin receptors. The inhibition can be demonstrated also when high concentrations of ferricyanide are used as electron acceptor. The possible mechanism behind the reported phenomena is discussed, and it is concluded that the transplasma membrane electron transport system can be involved in the process of cellular iron uptake.

Apoproteins↗
PubMed1987

The role of ascorbate in biomembrane energetics.

The mechanism(s) whereby membrane translocations are energized are poorly understood. Our work has focused on transmembrane microsomal and plasma membrane redox constituents as a means to energize membranes via alternative mechanisms complementary to ATP-driven processes. One such component is NADH-ascorbate free radical (mono- or semidehydroascorbate) oxidoreductase. This activity is associated with the trans or exit face of the Golgi apparatus, transport vesicles that move between the Golgi apparatus and the plasma membrane, and with the plasma membrane itself. Various lines of evidence, mostly indirect, link this activity to membrane translocations. Included is an apparent activation of the reductase in membranes when coated with clathrin, a single large polypeptide chain involved in exocytosis and in receptor-mediated and absorptive endocytosis. The results are consistent with a role of the ascorbate free radical as an acceptor for electron transport-mediated transfer of electrons from NADH perhaps to oxygen by coated membranes as a part of a mechanism to drive membrane translocations via generation of a proton gradient or of a membrane potential. Additionally, plasma membrane redox may be important in the regulation of cell growth, but a strict dependence on ascorbate free radical for the latter seems less likely than with internal endomembranes, where redox function may strictly depend upon the restricted pool of regeneratable acceptor that the ascorbate free radical provides.

Animals↗
PubMed1987

Diferric transferrin reductase in the plasma membrane is inhibited by adriamycin.

Diferric transferrin which is often necessary for growth of cells is reduced by the transplasma membrane electron transport system of HeLa cells with release of ferrous iron outside the cell. Reduction of external diferric transferrin is reflected in oxidation of internal NADH. Adriamycin, an antitumor drug, inhibits diferric transferrin reduction by the HeLa cells and inhibits concomittant oxidation of cytosolic NADH at concentrations, 10(-8)-10(-6)M, which inhibit cell growth. Isolated liver plasma membranes have an NADH diferric transferrin reductase activity which is inhibited by similar adriamycin concentrations. We propose that inhibition of cell growth by adriamycin can be based on inhibition of transplasmalemma diferric transferrin reductase.

Cell Membrane↗
PubMed1986

Redox reactions of tonoplast and plasma membranes isolated from soybean hypocotyls by free-flow electrophoresis.

Redox reactions were studied in more than 90% pure tonoplast and plasma membranes isolated by free-flow electrophoresis from soybean (Glycine max) hypocotyls. Both types of membrane contained a b-type cytochrome (alpha max = 561 nm) and a noncovalently bound flavin, two possible components of a transmembrane electron-transport chain. Isolated tonoplast and plasma membranes reduced ferricyanide, indophenol and various iron complexes with NADH or NADPH as electron donors. The redox activity was inhibited in tonoplast membranes by about 60% by 10 microM p-chloromercuribenzene sulfonate, 8% by 500 microM lanthanum nitrate and 10% by 100 microM nitrophenyl acetate. In contrast, the redox activity of isolated plasma membranes was inhibited by about 60% by 500 microM lanthanum nitrate or 100 microM nitrophenyl acetate, but only 25% by 10 microM p-chloromercuribenzene sulfonate. The results show that both tonoplast and plasma membranes of soybean contain active electron-transport systems, but that the two systems respond differently to inhibitors.

Cell Fractionation↗
PubMed1986

Transplasmalemma electron transport from cells is part of a diferric transferrin reductase system.

Intact cells are known to reduce external, impermeable electron acceptors. We now show that cells can reduce the iron in diferric transferrin at the cell surface and that this reduction reaction depends on the transferrin receptor as well as the transmembrane electron transport system. Reduction of external diferric transferrin is accompanied by oxidation of internal NADH which indicates that the transmembrane enzyme is an NADH diferric transferrin reductase. Highly purified liver plasma membranes have NADH diferric transferrin reductase activity which shows properties similar to the diferric transferrin reductases activity of intact cells. Cell growth stimulation by diferric transferrin and other impermeable oxidants which can react with the diferric transferrin reductase can be based on electron transport through he plasma membrane.

Cell Membrane↗
PubMed1986

Modification of transmembrane electron transport activity in plasma membranes of simian virus 40 transformed pineal cells.

Changes have been found in the plasma membrane enzyme system which carries out transmembrane electron transport and associated proton transport in Simian virus 40 (SV40) temperature-sensitive A (tsA) mutant-transformed rat pineal cell line, RPN209-1. This cell line was temperature-sensitive for the maintenance of transformation. RPN209-1 cells expressed the transformed phenotype (rapid growth, high cell density, and cloning in soft agar) at the permissive temperature (33 degrees C) and the nontransformed phenotype (slower growth, lower saturation density, and lower cloning efficiency in soft agar) at the nonpermissive temperature (40 degrees C). The reduction of external ferricyanide, hexaammine ruthenium and diferric transferrin was used to measure the transmembrane redox activity. The transformed RPN209-1 cells expressed a lower transmembrane redox activity, which is more sensitive to the antitumor drug adriamycin, when compared to the cells with a nontransformed phenotype. The lower transmembrane redox activity is associated with a decrease in the affinity for ferricyanide and a change in Vmax of the enzyme. Since the transformed cells have 25% lower concentration of NADH, the decrease in Vmax may be partly based on substrate limitation. Ionic strength variation in the assay media shows that the change in activity with transformation is not based on change in cell-surface change. Treatment with neuraminidase, however, indicates that sialic acid is important for enzyme activity, consistent with previous proposals that the transmembrane enzyme is a glycoprotein. The proton extrusion associated with transplasma membrane electron transport is increased in transformed cells relative to the rate of ferricyanide reduction. A relation between proton pumping transplasma membrane electron transport and growth stimulation by external oxidants is discussed.

Animals↗
PubMed1986

Decrease of NADH in HeLa cells in the presence of transferrin or ferricyanide.

The short-term incubation of HeLa cells in the presence of diferric transferrin or ferricyanide, which are reduced externally by the transplasma membrane reductase, produces a stoichiometric decrease in NADH and increase in NAD+, which is stimulated by insulin. The NADP/NADPH ratio does not change during 15 min incubation with the oxidants. The total pyridine nucleotide pool of HeLa cells is not affected. Incubation with apotransferrin and ferrocyanide, which cannot act as oxidants for transmembrane electron transport, does not change the pyridine nucleotide concentrations in the cells. Our results show that NADH can act as the internal electron donor for the reduction of external oxidants by the transmembrane reductase. It appears that oxidation of NADH by the transmembrane electron transport using ferricyanide or iron transferrin as external electron acceptors is sufficient to stimulate growth in HeLa cells.

Apoproteins↗
PubMed1986

Transplasmalemma electron transport is changed in simian virus 40 transformed liver cells.

Transplasma membrane electron transport activity by fetal rat liver cells (RLA209-15) infected with a temperature-sensitive strain of SV40 has been measured with cells grown at the restrictive temperature (40 degrees C) and permissive temperature (33 degrees C). The transformed cells grown at 33 degrees C had only one-half the rate of external ferricyanide reduction as the nontransformed cells held at 40 degrees C. Both the Km and Vmax for ferricyanide reduction were changed in the transformed state. The change in Vmax can be based on a decrease of NADH in the transformed cells. The change in rate with ferricyanide does not depend on change in surface charge. Reduction of external ferricyanide was accompanied by release of protons from the cells. The ratio of protons released to ferricyanide reduced was higher in the transformed cells than in the non-transformed cells. Since the transplasma membrane electron transport has been shown to stimulate cell growth under limiting serum, the changes in the plasma membrane electron transport and proton release in transformed cells may relate to modification of growth control.

Animals↗
PubMed1985

Oxidation of reduced pyridine nucleotides by plasma membranes of soybean hypocotyl.

Highly purified plasma membranes isolated from soybean hypocotyls by free-flow electrophoresis or by a two-phase polymer separation system oxidize reduced pyridine nucleotides, NADH or NADPH, at rates of 2-5 nanomoles/mg protein/min. These rates are not influenced by mitochondrial inhibitors or by inhibitors of the alternate respiratory pathway. The NADH oxidase has a Km of 200 microM NADH. The enzyme activity is stimulated by Ca2+ and Mg2+ ions. The function of this enzyme is unknown at present, but it may represent a redox-controlled proton pump linked to acidification.

Calcium↗
PubMed1985

Bleomycin control of transplasma membrane redox activity and proton movement in HeLa cells.

Bleomycin, tallysomycin A, tallysomycin S10b and copper-bleomycin have been tested for their capacity to inhibit the transplasma membrane electron transport and associated proton release by HeLa cells. Transplasma membrane redox activity is measured using reduction of external ferricyanide by the cells. At 75 micrograms/ml bleomycin, tallysomycin A and tallysomycin S10b gave a maximum of 65% inhibition of the ferricyanide reduction rate; half-maximum inhibition was observed at 30 micrograms/ml. The copper-bleomycin complex was slightly more effective as an inhibitor with half-maximum inhibition at 20 micrograms/ml. Survival of cells after 1 hr of drug treatment was 50% at 25 micrograms/ml for bleomycin and copper-bleomycin and at 75 micrograms/ml for tallysomycin A. Tallysomycin A and tallysomycin S10b gave 75 to 83% inhibition of ferricyanide-induced proton extrusion, respectively at 50 micrograms/ml, whereas bleomycin and copper-bleomycin appeared to be slightly less effective with 50 to 60% inhibition, respectively, at 50 micrograms/ml. In all aspects studied, which included transplasma membrane ferricyanide reduction, ferricyanide-induced proton release, and cell survival, there were significant effects by these compounds on HeLa cells in the range of 25-50 micrograms/ml.

Bleomycin↗
PubMed1985

alpha-Adrenergic stimulation of trans-sarcolemma electron efflux in perfused rat heart. Possible regulation of Ca2+-channels by a sarcolemma redox system.

The role of trans-sarcolemma membrane electron efflux in the alpha-adrenergic control of Ca2+ influx in perfused rat heart was examined. Electron efflux was measured by monitoring the rate of reduction of extracellular ferricyanide and compared with changes in contractility, as an indirect assessment of changes in cytoplasmic Ca2+ concentration. Methoxamine and phenylephrine each increased the rate of ferricyanide reduction from 80 to approx. 114 nmol/min per g wet wt. of heart, with half-maximal activation occurring at 10 microM for each agonist. Activation of the rate of ferricyanide reduction by both 10 microM methoxamine and 10 microM phenylephrine was blocked by the alpha-adrenergic antagonist, phenoxybenzamine, but not by the beta-antagonist, propranolol. Stimulation of the rate of ferricyanide reduction by the alpha-agonist coincided with the increase in contractility, each reaching maximum values at approx. 80 s. Removal of the alpha-agonists led to parallel decreases in contractility and the rate of reduction, each returning to pre-stimulation values in approx. 400 s. In addition, the relationship between Ca2+ and ferricyanide reduction was examined. Perfusion of the heart with medium containing 6 mM CaCl2 significantly increased contractility and the rate of ferricyanide reduction. Perfusion of the heart with low Ca2+ diminished contractility, did not affect the rate of ferricyanide reduction, but amplified the stimulatory effect of methoxamine on this rate. The increase in ferricyanide reduction by alpha-adrenergic agonists resulted from a change in the apparent Vmax, indicative of an increase in electron efflux sites in the plasma membrane. It is concluded that alpha-adrenergic control of electron efflux closely parallels changes in contractility and therefore changes in the cytoplasmic concentration of Ca2+. The data suggest that alpha-agonist-mediated changes in electron efflux may lead to Ca2+ influx.

Adrenergic alpha-Agonists↗
PubMed1985

Evidence for Ca++-calmodulin control of transplasmalemma electron transport in carrot cells.

Cultured carrot cells exhibit transmembrane ferricyanide reduction through a plasma membrane redox system, which may be associated with an iron reduction and uptake system in plant roots. Here we provide evidence for the inhibition of transplasma membrane ferricyanide reduction by four different Ca2+-calmodulin type antagonists, calmidazolium, trifluoperazine, pimozide and fluphenazine. These compounds inhibit in low concentrations (approximately 5-10 microM) in a time-dependent manner. Higher concentrations (50-100 microM) are required to inhibit transmembrane ferricyanide reduction in 10 min rather than in 30 min. The permeable calcium chelator, TMB-8, also inhibits transmembrane ferricyanide reduction in carrot cells. Since the redox system is controlled by hormones, the effects of anticalmodulin agents on hormone response may be mediated through the redox system.

Calcium↗
PubMed1985

Transmembrane ferricyanide reduction in carrot cells.

Carrot cells (Daucus carota) grown in tissue culture are capable of reducing the non-permeable electron acceptor, ferricyanide, with concomitant proton extrusion from the cell. Optimum conditions for transmembrane ferricyanide reduction include a pH of 7.0-7.5 in a medium containing 10 mM each KCl, NaCl and CaCl2. Data are shown to prove that transmembrane ferricyanide reduction is an enzymatic process. It does not depend on the secretion of phenolics from the cell within the time limits of the assay (10 min). The presence of broken cells and cell fragments are excluded on the basis of stimulation or only slight inhibition by mitochondrial inhibitors. However, transmembrane ferricyanide reduction by carrot cells is inhibited about 50% by various glycolysis inhibitors, which are presumed to reduce the internal levels of NADH. Treatment of cells with p-diazoniumbenzenesulfonic acid, a non-permeant membrane modifying agent, also inhibits transmembrane ferricyanide reduction more than 90%. The data presented support the existence of a transplasma membrane redox system in carrot cells.

Cell Membrane↗
PubMed1985

Transmembrane redox in control of cell growth. Stimulation of HeLa cell growth by ferricyanide and insulin.

The impermeable electron acceptor ferricyanide stimulates the growth of HeLa cells in the absence of serum and increases cell replication with limiting amounts of serum (0.75%). Maximum growth stimulation occurs at low ferricyanide concentration from 0.01 to 0.1 mM. Higher ferricyanide concentrations inhibit growth on serum. Addition of insulin enhances the stimulating effect of ferricyanide. Increase in the transplasmalemma electron transport activity in the presence of insulin is also observed by measuring the rate of ferricyanide reduction by cells. There is a close correlation between insulin stimulation of ferricyanide reduction and insulin induction of cell proliferation and attachment. In addition to ferricyanide, the growth response is observed with other impermeable oxidants, such as indigotetrasulfonate and hexaamine ruthenium III, which are reduced by the transplasma membrane electron transport system. Inactive oxidants such as cytochrome c do not stimulate cell growth. Ferrocyanide does not stimulate growth. We propose that electron flow through the transplasma membrane electron transport system stimulates growth and that insulin acts to increase that flow.

Blood↗