Plasma Membrane Redox Reactions Research Articles

Chromium (VI) induced changes in growth and root plasma membrane redox activities in pea plants.

The effect of chromium (Cr) on growth as well as root plasma membrane redox reactions and superoxide radical production was studied in pea (Pisum sativum L. cv. Azad) plants exposed for 7 days to 20 and 200 microM Cr (VI), respectively, supplied as potassium dichromate. The growth of pea plants declined significantly at 200 microM Cr, as indicated by reduced leaf area and biomass. Relative to the control plants (no Cr exposure), the Cr content of roots increased significantly, both at 20 and 200 microM Cr. Following exposure to 200 microM Cr, there was a significant increase in root lipid peroxidation and hydrogen peroxide (H(2)O(2)) content, while both the Fv/Fm ratio and chlorophyll content were reduced. Exposure to Cr increased NADPH-dependent superoxide production in pea root plasma membrane vesicles, with the effect being more significant at 200 microM Cr than at 20 microM Cr. Treatment with Cr rapidly increased the activities of NADPH oxidase: relative to the controls, plants exposed to 20 microM Cr showed approximately a 67% increase in activity while there was a threefold increase in those plants exposed to 200 microM Cr. NADH-ferricyanide oxido-reductase activity was found to be inhibited by 16 and 51% at 20 and 200 microM Cr, respectively. The results of this study suggest that exposure to excess Cr damages pea root plasma membrane structure and function, resulting in decreased photosynthesis and poor plant growth.

The plasma membrane redox system in human platelet functions and platelet-leukocyte interactions

The plasma membrane electron transport is crucial for blood coagulation and thrombosis, since reactive oxygen species and thiol changes, generated by plasma membrane redox reactions, modulate activation of platelets, as well as their interaction with leukocytes. Several antioxidants are linked to this system; thus, platelets are also able to counterbalance radical production and to regulate thrombus growth. Aim of this review is to give an update on the plasma membrane redox system in platelets, as well as on its role in platelet functions and leukocyte-platelet cross-talk.

Relationship between IAA-induced elongation growth and plasma membrane NADH oxidase in soybean (Glycine max Merr.) hypocotyls

A relationship between the activity of NADH oxidase of the plasma membrane and the IAA-induced elongation growth of hypocotyl segments in etiolated soybean (Glycine max Merr.) seedlings was investigated. The plasma membrane NADH oxidase activity increased in parallel to IAA effect on elongation growth in hypocotyl segments. Actually, NADH oxidase activity was stimulated 3-fold by 1 u,M IAA, and the elongation rate of segments was stimulated 10-fold by 10 iM IAA. The short-term elongation growth kinetics, however, showed that the IAA-induced elongation of hypocotyl segments was completely inhibited by plasma membrane redox inhibitors such as actinomycin D and adriamycin, at 80 μM and 50 μM respectively. In addition, 1 mM actinomycin D inhibited the IAA-stimulated NADH oxidase activity by about 80%. However, adriamycin had no effect on NADH oxidase activity of plasma membrane vesicles. Based on these results, the plasma membrane redox reactions seemed to be involved in IAA-induced elongation growth of hypocotyls, and the redox component responding to IAA was suggested to be NADH oxidase.

The NADH oxidizing system of the plasma membrane and metabolic signal control

The development of ideas concerning plasma membrane redox reactions in normal and transformed animal cells is described, with emphasis on transferrin and ceruloplasmin. Control by hormones and growth factors, as well as the NAD+/NADH ratio in the cell are important in distinguishing the two types of cells.

Involvement of ascorbic acid and ab-type cytochrome in plant plasma membrane redox reactions

Higher plant plasma membranes contain ab-type cytochrome that is rapidly reduced by ascorbic acid. The affinity towards ascorbate is 0.37 mM and is very similar to that of the chromaffin granule cytochromeb 561. High levels of cytochromeb reduction are reached when ascorbic acid is added either on the cytoplasmic or cell wall side of purified plasma membrane vesicles. This result points to a transmembrane organisation of the heme protein or alternatively indicates the presence of an effective ascorbate transport system. Plasma membrane vesicles loaded by ascorbic acid are capable of reducing extravesicular ferricyanide. Addition of ascorbate oxidase or washing of the vesicles does not eliminate this reaction, indicating the involvement of the intravesicular electron donor. Absorbance changes of the cytochromeb α-band suggest the electron transfer is mediated by this redox component. Electron transport to ferricyanide also results in the generation of a membrane potential gradient as was demonstrated by using the charge-sensitive optical probe oxonol VI. Addition of ascorbate oxidase and ascorbate to the vesicles loaded with ascorbate results in the oxidation and subsequent re-reduction of the cytochromeb. It is therefore suggested that ascorbate free radical (AFR) could potentially act as an electron acceptor to the cytochrome-mediated electron transport reaction. A working model on the action of the cytochrome as an electron carrier between cytoplasmic and apoplastic ascorbate is discussed.

Plant Defense Response to Fungal Pathogens (II. G-Protein-Mediated Changes in Host Plasma Membrane Redox Reactions).

Elicitor preparations containing the avr5 gene products from races 4 and 2.3 of Cladosporium fulvum, and tomato (Lycopersicon esculentum L.) cells containing the resistance gene Cf5 were used to investigate the involvement of redox processes in the production of active oxygen species associated with the plant response to the fungal elicitors. Here we demonstrate that certain race-specific elicitors of C. fulvum induced an increase in ferricyanide reduction in enriched plasma membrane fractions of tomato cells. The addition of elicitors to plasma membranes also induced increases in NADH oxidase and NADH-dependent cytochrome c reductase activities, whereas ascorbate peroxidase activity was decreased. These results suggest that changes in the host plasma membrane redox processes, transferring electrons from reducing agents to oxygen, could be involved in the increased production of active oxygen species by the race-specific elicitors. Our results also show that the dephosphorylation of enzymes involved in redox reactions is responsible for the race-specific induced redox activity. The effects of guanidine nucleotide analogs and mastoparan on the activation of plasma membrane redox reactions support the role of GTP-binding proteins in the transduction of signals leading to the activation of the defense response mechanisms of tomato against fungal pathogens.

The possible role of redox-associated protons in growth of plant cells.

The protons excreted by plant cells may arise by two different mechanisms: (1) by the action of the plasma membrane H(+)-ATPase and (2) by plasma membrane redox reactions. The exact proportion from each source is not known, but the plasma membrane H(+)-ATPase is, by far, the major contributor to proton efflux. There is still some questions of whether the redox-associated protons produced by NADH oxidation on the inner side of the plasma membrane traverse the membrane in a 1:1 relationship with electrons generated in the redox reactions. Membrane depolarization observed in the presence of ferricyanide reduction by plasma membranes of whole cells or tissues or the lag period between ferricyanide reduction and medium acidification argue that only scalar protons may be involved. The other major argument against tight coupling between protons and electrons involves the concept of strong charge compensation. When ferricyanide is reduced to ferrocyanide on the outside of cells or tissues, an extra negative charge arises, which is compensated for by the release of H+ or K+, so that the total ratio of increased H+ plus K+ equals the electrons transferred by transmembrane electron transport. These are strong arguments against a tight coupling between electrons and protons excreted by the plasma membrane. On the other hand, there is no question that inhibitor studies provide evidence for two mechanisms of proton generation by plasma membranes. When the H(+)-ATPase activity is totally inhibited, the addition of ferricyanide induces a burst of extra proton excretion, or vice versa, when plasma membrane redox reactions are inhibited, the H(+)-ATPase can function normally. Since plasma membrane redox reactions and associated H+ excretion are related to growth it is possible that in plants the ATPase-generated protons have a different function from redox-associated protons. The H(+)-ATPase-generated protons have been considered for many years to be necessary for cell wall expansion, allowing elongation to take place. A special function of the redox-generated protons may be in initiating proliferative cell growth, based on the presence of a hormone-stimulated NADH oxidase in membranes of soybean hypocotyls and stimulation of root growth by low concentrations of oxidants. Here we propose that this NADH oxidase and the redox protons released by its action control growth. The mechanism for this may be the evolution of protons into a special membrane domain, from which a signal to initiate cell proliferation may originate, independent of the action of the H(+)-ATPase-generated protons.(ABSTRACT TRUNCATED AT 400 WORDS)

The effect of selected inhibitors on plasma membrane redox reactions and proton excretion by carrot cells

Five different calmodulin antagonists, calmidazolium, trifluoperazine, chloroproazine, W-7 and W-5, were tested for inhibition of plasma membrane redox reactions on cultures carrot cells. Their effects varied. NADH oxidation was inhibited about 50% by calmidazolium and W-7, while the inactive form, W-5, stimulated NADH oxidation. Proton excretion by cultured carrot cells was totally inhibited by 40 μM calmidazolium, but it was differentially inhibited by trifluoperazine. Twenty micromolar trifluoperazine inhibited the basal rate of proton excretion 100%; 50 μM trifluoperazine inhibited about 90% of the redox-associated proton excretion. Inhibition of plasma membrane redox reactions and associated proton excretion by a few selected calmodulin antagonists may point to the association of Ca 2+-calmodulin as second messengers leading to cell growth.

The effect of inhibitors of plasma membrane redox reactions on proton excretion by plant cells

Plant cells excrete protons via an electrogenic proton pump, the K+‐stimulated, Mg2+‐dependent H+‐ATPasc, located on the cytoplasmic side of the plasma membrane. Plasma membrane redox reactions are also coupled to proton excretion. Various inhibitors were used on carrot (Daucus carola L.) cells in an attempt to distinguish between the two processes. Inhibitors of electron transport reactions in the plasma membrane (chloroquine, 8‐hydroxyquinolinc, 4,7‐dichloroquinolinc and retinoic acid) inhibited ferricyanide‐induced proton excretion by 37–100%, while they inhibited potassium ferricyanide reduction, a measure of plasma membrane redox activity, by 42–100%. The above‐mentioned quinolines and retinoic acid inhibited cell growth by 49–98%, with the exception of chloroquine, which stimulated carrot cell growth by 36%.

Inhibition of plasma membrane redox activities and elongation growth of soybean

NADH‐ferricyanide oxido‐reductase (EC 1,6,99,3) of purified plasma membrane vesicles isolated by aqueous two‐phase partition from segments of etiolated soybean [Glycine max (L.) Merr. cv. Williams] hypocotyls was used as a measure of plasma membrane redox activity. Elongation growth of hypocotyl segments floated on the solutions was determined in parallel. Cis‐platinum (II) diammine dichloride (cis‐platin), adriamycin and p‐nitrophenylacetate, agents known to inhibit cell proliferation and plasma membrane redox activities in mammalian cells inhibited both NADH‐ferricyanide oxido‐reductase of the isolated membrane vesicles and elongation growth of intact hypocotyl segments. Auxin(2,4‐dichlorophenoxyacetic acid)‐induced growth of the isolated segments was inhibited preferentially at drug concentrations where control growth was affected only slightly. The findings suggest a connection between plasma membrane redox reactions and the control of elongation growth in plants.

Use of an electrochemical technique to study plasmamembrane redox reactions in cultured cells of Daucus carota L.

By exploiting the electron transfer reactions of ferricyanide-ferrocyanide at a gold electrode, an electrochemical technique was devised and successfully employed for the measurement of extracellular ferricyanide reduction or ferrocyanide oxidation by carrot (Daucus carota L.) cells grown in suspension culture. This technique eliminated problems of cell damage and insensitivity encountered when these activities are measured spectrophotometrically in magnetically stirred cell suspensions. Cells harvested from the mid-exponential phase of culture growth catalysed a rapid reduction of ferricyanide which was accompanied by H(+) extrusion and was stimulated by ethanol. These cells also oxidised ferrocyanide, which was not associated with H(+) extrusion. These observations can be explained on the basis of a plasmamembrane located, H(+)-translocating redox system. The electrochemical technique is a useful method of studying this system.

Inhibition of plasma membrane NADH dehydrogenase by adriamycin and related anthracycline antibiotics.

Doxorubicin (adriamycin) is cytotoxic to cells, but the biochemical basis for this effect is unknown, although intercalation with DNA has been proposed This study suggests that the cytotoxicity of this drug may be due to inhibition of the plasma membrane redox system, which is involved in the control of cellular growth. Concentrations between 10(-6) - 10(-7) M adriamycin inhibit plasma membrane redox reactions greater than 50%. AD32, a form of adriamycin which does not intercalate with DNA, but is cytotoxic, also inhibits the plasma membrane redox system. Thus, the cytotoxic effects of adriamycin, which limit its use as a drug, may be based on the inhibition of a transplasma membrane dehydrogenase involved in a plasma membrane redox system.