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Biomedical subjects

H Sentenac

Publications and source records attributed to H Sentenac.

29 records · Page 2Linked to original sources
PubMed1993

A test for screening monoclonal antibodies to membrane proteins based on their ability to inhibit protein reconstitution into vesicles.

The hypothesis that the binding of an antibody to a membrane protein is likely to prevent the reconstitution of the protein into liposomes was checked, by using the plant plasma membrane H(+)-ATPase (EC 3.6.1.35) as a model system, and two reconstitution procedures: spontaneous insertion (SI) of purified H(+)-ATPase into preformed liposomes, and a detergent-mediated reconstitution (DMR) procedure allowing the reconstitution of the whole membrane protein content. Nine monoclonal antibodies (MABs) raised against H(+)-ATPase were tested. None affected the functioning of the enzyme reconstituted in liposomes, suggesting that the probability to obtain an inhibitory MAB is low. Five MABs inhibited its SI, and seven inhibited its reconstitution in the DMR procedure. These results indicate that it is possible to screen antibodies directed against membrane protein, by making use of their ability to inhibit the reconstitution of these proteins.

Antibodies, Monoclonal↗
PubMed1992

Cloning and expression in yeast of a plant potassium ion transport system.

A membrane polypeptide involved in K+ transport in a higher plant was cloned by complementation of a yeast mutant defective in K+ uptake with a complementary DNA library from Arabidopsis thaliana. A 2.65-kilobase complementary DNA conferred ability to grow on media with K+ concentration in the micromolar range and to absorb K+ (or 86Rb+) at rates similar to those in wild-type yeast. The predicted amino acid sequence (838 amino acids) has three domains: a channel-forming region homologous to animal K+ channels, a cyclic nucleotide-binding site, and an ankyrin-like region.

Amino Acid Sequence↗
PubMed1988

H Cotransports in Corn Roots as Related to the Surface pH Shift Induced by Active H Excretion.

The surface pH shift induced by active H(+) excretion in corn (Zea mays L.) roots was estimated using acetic acid influx as a pH probe (H Sentenac, C Grignon 1987 Plant Physiol 84: 1367-1372). At constant bulk pH, buffering the medium strongly reduced the magnitude of the surface pH shift. This was used to study the effect of surface pH shift on H(+) cotransports. In the absence of buffers, the surface pH shift increased with the bulk pH. Buffers decreased (32)Pi influx and this effect was stronger at pH 7.2 than at pH 5.8, and stronger in the absence than in the presence of an inhibitor of the proton pump (vanadate). Buffers exerted a similar depressive and pH-dependent effect on net NO(3) (-) uptake. They hyperpolarized the cell membrane, and stimulated (86)Rb(+) influx, K(+):H(+) net exchange, and malate accumulation. These results are consistent with the hypothesis that H(+) accumulation at the cell surface is effective in driving H(+) reentry. We concluded that the surface pH shift due to proton pump activity is involved in the energetic coupling of H(+) cotransports.

Journal Article↗
PubMed1987

Effect of H Excretion on the Surface pH of Corn Root Cells Evaluated by Using Weak Acid Influx as a pH Probe.

The initial rate of (2-(14)C)acetic acid (AA) uptake by corn roots was used for probing the dependency of the root cell surface pH on H(+) excretion. AA influx was linearly related to AA concentration, dependent on the concentration of the undissociated form (AH), unaffected by variations of the membrane potential, and was thus assumed to result mainly from the free diffusion of AH across the membrane. Various treatments (vanadate, dicyclohexylcarbodiimide, hypoxia, nitrate, root ageing, fusicoccin) were used to vary H(+) flux while the medium pH was maintained constant. There was a positive relation between AA influx and the net H(+) efflux. This relation disappeared when the proton buffering strength of the absorption medium was increased. These results indicate that the pH at the membrane surface was lowered by H(+) excretion, even in situations where the bulk (pH) was unaffected. The depressive effect of vanadate on AA influx was counteracted by acidifying the medium in order to estimate this pH shift: -1.2 pH units in 12.5 millimolar K(2)SO(4) (pH 6.8). Substituting AA by butyric acid showed that this estimation was not dependent of the probe used.

Journal Article↗
PubMed1985

Effect of pH on Orthophosphate Uptake by Corn Roots.

Orthophosphate (Pi) influx in washed corn roots was studied with experimental conditions allowing a distinction of pH effects on Pi ionization in the medium and on the transport system itself. There appeared to be no relationship between the pH dependencies of membrane potential, H(+) secretion, and (32)Pi influx. The Pi uptake versus pH curves were compared to the calculated ones describing the concentrations of the different ionized Pi forms in the medium and in the cell walls; the latter were obtained using the theoretical model described by Sentenac and Grignon (1981) Plant Physiol 68: 415-419). The conclusion was that the transported form is H(2)PO(4) (-) and the concentration sensed by the transport system is the local one. The ionic compositions of experimental media were manipulated to ensure constant pH and various H(2)PO(4) (-) concentrations, or constant H(2)PO(4) (-) concentration and various pH values in the walls. The kinetic analysis of the results in the micromolar range showed that the transport system has an intrinsic sensitivity to pH, and is switched from a low activity state at pH > 6 to a high activity one at pH < 4 (pH in the walls). This change could be triggered by the protonation of a group with pK 5.5.

Journal Article↗
PubMed1984

Effects of physostigmine, paraoxon and soman on brain GABA level and metabolism.

The effects of sublethal doses of physostigmine, paraoxon and soman on GABA levels and metabolism were studied in small rat brain areas (hypothalamus, striatum, cerebellum, rest of the brain). Physostigmine induced a significant decrease in striatal GABA level and a reduction of synthesis index in the rest of the brain while organophosphates have little or no effect on GABA level and metabolism. This work provides new data about the physostigmine effect on brain GABA which could be related to the action of the anticholinesterase agents on other non-cholinergic neurotransmitters.

4-Aminobutyrate Transaminase↗
PubMed1983

Brain catecholamine metabolism changes and hypothermia in intoxication by anticholinesterase agents.

Sublethal doses of physostigmine, paraoxon and soman induce a short-lasting fall in rat core temperature potentiated by alpha-methyl-para-tyrosine (alpha-MT) (early effects). When the own hypothermic effect of the anticholinesterase agent has disappeared (late effects), alpha-MT induces a new decrease in temperature. Parallel biochemical studies of catecholamine levels and turnover were performed in several brain areas. The norepinephrine (NE) turnover is generally increased particularly in the hypothalamus, suggesting that NE hypothalamic changes might be linked to a latent perturbation of thermoregulatory mechanisms. Furthermore, it was shown that soman acts differently from the other drugs by inducing quite important changes in both NE and dopamine levels.

Animals↗
PubMed1981

A model for predicting ionic equilibrium concentrations in cell walls.

Purified cell walls were prepared from roots of Horse bean (Vicia faba L., var. minor) and Yellow Lupine (Lupinus luteus L.). Two methods were used: (a) grinding and (b) lysis of the endocellular contents by Triton X-100. The Ca(2+), Mg(2+), and K(+) contents were determined after incubation in various solutions in such a manner that the measurements gave access to the undisturbed equilibrium contents. The results were used to test a model which describes the ionic atmosphere in the cell walls as a result of simultaneous electrostatic interactions between free ions (Donnan effect) and specific association equilibria, including acido-basic ones. This model correctly accounted for the whole set of experimental results and predicted the values of the unmeasurable local concentrations and pH.

Journal Article↗
PubMed1980

Effect of tabernanthine on the turnover time of brain catecholamines in normal and hypobaric hypoxic rats.

The central effects of tabernanthine on noradrenaline and dopamine turnover times were studied in the hypothalamus, the striatum and the remainder of the brain of normal and hypobaric hypoxic rats, the latter state corresponding to 5,200m (410mm Hg) and 7,000m (320mm Hg). Catecholamine cerebral level were not modified by the drug in either instance. At normal atmospheric pressure the catecholamine turnover times were slightly decreased by tabernanthine. Hypobaric hypoxia alone increased noradrenaline and dopamine turnover times by inhibiting oxygen-dependent enzymes. Tabernanthine antagonized the effect of hypobaric hypoxia especially in dopaminergic areas. Antagonism was complete at 5,200m but only partial at 7,000m. This phenomenon may be related to the stimulatory properties of the drug.

Alkaloids↗