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D Bataille

Publications and source records attributed to D Bataille.

At least 37 records · Page 2Linked to original sources
PubMed1993

GEP31, a new gastric epithelial protein, early expressed during ontogenesis.

A set of monoclonal antibodies directed against various gastric markers was produced in order to study the developmental expression of the gastric mucosa. A previously described monoclonal antibody, mab 146.14, labeled the proton pump (H+, K+) ATPase specifically located in gastric parietal cells. Mabs 15.1 and 121.17 revealed the mucus of mucous neck cells and mucous surface cells, respectively. By addition, mab 81.1 was directed against a cell surface membrane protein antigen composed of a major 31 kDa component (called GEP31). Our study mainly focused on the characterization of GEP31. This protein was typically located in the gastrointestinal epithelial tract (stomach, small intestine, colon). Moreover, interesting features were observed during the study of the early ontogenesis of the gastric mucosa. The 31 kDa protein was detected at the onset of gland formation (day 18), and a gradual increase in expression of the protein could be observed between day 18 and day 19. Furthermore, a comparative study of the expression of different terminal differentiation markers of gastric epithelial cells ((H+, K+) ATPase, mucins) during the early period of ontogenesis revealed that GEP31 could be detected well before the appearance of these markers. To our knowledge, GEP31 thus appears as the earliest expressed specific cell surface epithelial gastric antigen described to date. Furthermore, the partial N-terminal amino acid sequence of GEP31 was determined and revealed that it is not a known protein.

Amino Acid Sequence↗
PubMed1992

Endosulfine, an endogenous peptidic ligand for the sulfonylurea receptor: purification and partial characterization from ovine brain.

Antidiabetic sulfonylureas act through receptors coupled to ATP-dependent potassium channels. Using the binding of [3H]glibenclamide, a highly potent sulfonylurea, to rat brain membranes to follow the purification procedure, we extracted from ovine brain, purified, and partially characterized two peptides that are endogenous ligands for the central nervous system sulfonylurea receptors. These peptides, referred to as alpha and beta endosulfine, differ by their isoelectric points, the beta form being more basic. Each form of endosulfine is recognized equally by the sulfonylurea receptors from the central nervous system and from insulin-secreting beta cells. In the same concentration range that is active on the receptors, beta endosulfine releases insulin from a beta-cell line. Endosulfine is a good candidate for being implicated in the physiology of beta cells and their disorders (e.g., type II diabetes) and in certain pathologies related to modifications of ion fluxes.

ATP-Binding Cassette Transporters↗
PubMed1992

Role of G protein beta gamma subunits in the regulation of the plasma membrane Ca2+ pump.

In Zajdela hepatoma cells (ZHC) the plasma membrane Ca2+ pump displayed no sensitivity to glucagon (19-29) (mini-glucagon), whereas in hepatocyte this metabolite of glucagon evoked a biphasic regulation of the Ca2+ pump system via a cholera toxin-sensitive G protein. Analysis of G protein subunits in ZHC membranes indicated the presence of cholera toxin-sensitive Gs alpha and G beta gamma proteins, whose functionality was manifested by GTP and NaF stimulation of adenylylcyclase activity, and pertussis toxin-catalyzed ADP-ribosylation of Gi alpha, respectively. However, immunoblotting experiments suggested a lower content in beta gamma subunits in ZHC as compared with hepatocyte plasma membranes. Complementation of ZHC or hepatocyte plasma membranes with purified beta gamma subunits from transducin (T beta gamma) caused inhibition of the basal activity of the Ca2+ pump at 10 and 300 ng/ml, respectively, and revealed (in ZHC) or increased (in hepatocytes) sensitivity of the system to mini-glucagon. After cholera toxin treatment of ZHC, T beta gamma no longer reconstituted the response of the Ca2+ pump to mini-glucagon, suggesting that the mechanism of beta gamma action is dependent on an association with the alpha subunit of a cholera toxin-sensitive G protein. It is concluded that G beta gamma subunits control both the basal activity of the plasma membrane Ca2+ pump and its inhibition by mini-glucagon.

Adenosine Diphosphate Ribose↗
PubMed1992

N-acetyl oxyntomodulin30-37: pharmacokinetics and activity on gastric acid secretion.

Oxyntomodulin, an intestinal hormone which inhibits gastric acid secretion, is composed of glucagon and a C-terminal octapeptide. This octapeptide mimics the biological activity of the hormone. We have studied the activity of the N-acetyl octapeptide, partially protected against enzymatic degradation, on pentagastrin-, histamine- and milk meal-stimulated secretion in conscious rats and compared it to that of oxyntomodulin and its derivatives. The N-acetylated octapeptide had a 3-fold higher potency than the octapeptide on pentagastrin-stimulated secretion. On histamine-stimulated secretion, the differences between the acetylated and the free octapeptides were that the former displayed a dose-response curve parallel to that of oxyntomodulin and a 4-fold higher potency. The increase in potency appears to be related in part to a decrease in the metabolic clearance rate in vivo (6-fold) and, in vitro, to an increase in half-life (3-fold) when incubated with rat liver plasma membranes. Similarly to the free octapeptide, the acetylated form decreased acid secretion stimulated by a milk meal, when infused before the meal. Acetylation of the Lysine side chains resulted in a totally inactive molecule. The results indicate that acetylating the N-terminus of the octapeptide of oxyntomodulin increases the similarities with the natural hormone.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylation↗
PubMed1992

Characterization of binding sites for oxyntomodulin on a somatostatin-secreting cell line (RIN T3).

Oxyntomodulin (OXM), a glucagon-containing peptide extended at its C-terminal end by an octapeptide, is a potent inhibitor of gastric acid secretion in rat and man. OXM appears to act on gastric mucosa at least partially through a stimulation of gastric somatostatin release. We have investigated the effects of OXM on a somatostatin-secreting cell line (RIN T3) derived from a radiation-induced rat insulinoma and characterized specific binding sites for this peptide. OXM increased somatostatin release with an ED50 of 2.3 nM. OXM also stimulated the cAMP accumulation in intact RIN T3 cells and adenylate cyclase activity in RIN T3 cell membranes with ED50 values of 0.5 and 11 nM, respectively. On these parameters, glucagon was 10-30 times less potent than OXM. Forskolin, isobutylmethylxanthine, and 8-bromo-cAMP mimicked the effect of OXM on somatostatin release. Specific binding for mono-[125I]OXM was dependent upon time and membrane concentration. Binding of mono-[125I]OXM was inhibited by OXM and glucagon in a concentration-dependent manner, with dissociation constants (Kd) of 4.5 and 43 nM, respectively. The nonhydrolyzable analogs of GTP (guanosine 5',3-O-(thio)triphosphate and guanosine 5' (beta,gamma-imino)triphosphate decreased the binding of mono-[125I]OXM to its binding sites. Covalent cross-linking of mono-[125I]OXM or mono-[125I]glucagon to RIN T3 cell membranes followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated a single radiolabeled band at 63,000 mol wt, which differed from that observed after cross-linking with liver plasma membranes (55,000 mol wt). These results demonstrate the presence of specific high affinity binding sites for OXM in a somatostatin-secreting cell line (RIN T3) and their coupling to adenylate cyclase via guanine nucleotide-binding proteins.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Oxyntomodulin-like immunoreactivity: diurnal profile of a new potential enterogastrone.

The biological specificity of oxyntomodulin toward the gastric mucosa results from its C-terminal octapeptide. A RIA using a specific antibody raised against this region permitted quantification of the whole set of proglucagon-derived peptides that interact with the oxyntomodulin recognition systems, corresponding to the new concept of oxyntomodulin-like-immunoreactivity (OLI). The present report describes the physiological 24-h OLI profile in human plasma (eight men and eight women; mean age, 45 yr; range, 20-77 yr). Blood was withdrawn every hour from 0700-1900 h and every 2 h from 2100-0500 h. A meal-dependent profile was found for circulating OLI, with basal values (60 +/- 7 ng/L) at 0500 h and rises elicited by each food intake. The highest value (136 +/- 21 ng/L) was obtained at 2100 h. Plasma concentrations and diurnal variations of OLI were similar to those of the other intestinal peptides known to exert an endocrine function. The mean circulating OLI values increased with age, whereas no change was noticed according to sex. The inhibitory effect exerted by the peptides of the OLI family on gastric acid secretion, the meal dependence of their plasma concentrations, and the observed synchronism of their diurnal profile with that previously described for somatostatin make them candidates for an enterogastrone action.

Age Factors↗
PubMed1991

Oxyntomodulin and related peptides control somatostatin secretion in RIN T3 cells.

We studied the effects of oxyntomodulin (OXM), of its C-terminal (19-37) fragment (OXM (19-37)) and of glucagon (GLU) on somatostatin release, cyclic AMP accumulation and inositol phosphate turnover in somatostatin-secreting RIN T3 cells in culture. Rapid changes in cellular free Ca2+ were also measured using fura-2. Carbachol was used as a control test agent for the parameters involving the inositol phosphate/Ca2+ cascade. OXM, GLU and OXM (19-37) were all able to stimulate somatostatin release with relative ED50 of approx. 1, 22 and 45, respectively. OXM and GLU stimulated cyclic AMP levels with relative ED50 of approx. 1 and 30, respectively, whereas OXM (19-37) was totally ineffective on this parameter. In contrast to carbachol, none of the peptides significantly modified the inositol phosphate turnover or induced rapid changes in cellular free Ca2+. We conclude that the RIN T3 cells contain a receptor-cyclic AMP system similar to that found in gastric mucosa and that this system is linked to somatostatin release. Another receptor-second messenger mechanism linked to somatostatin release is triggered by the (19-37) fragment. This mechanism is not the inositol phosphate/Ca2+ cascade triggered in the same cells by cholinergic agents.

Animals↗
PubMed1991

Oxyntomodulin and its (19-37) and (30-37) fragments inhibit histamine-stimulated gastric acid secretion in the conscious rat.

Oxyntomodulin, a hormone released from jejuno-ileum and composed of the glucagon sequence extended by a C-terminal octapeptide displays original tissue specificity for the gastric mucosa. The aim of this study was to compare the effect of oxyntomodulin on histamine (0.4 mg/kg per h)-stimulated gastric acid secretion in the conscious rat to that of three of its fragments: oxyntomodulin-(19-37) produced from oxyntomodulin by enzymatic cleavage, oxyntomodulin-(30-37) corresponding to the molecular difference between oxyntomodulin and glucagon and oxyntomodulin-(32-37) produced during proglucagon processing to glucagon. The metabolic clearance rate (MCR) in the anesthetized rat was evaluated for each peptide. Oxyntomodulin, oxyntomodulin-(19-37) and oxyntomodulin-(30-37) inhibited the histamine-stimulated gastric acid secretion dose dependently. The dose-response curves for oxyntomodulin and oxyntomodulin-(19-37) were parallel. The potency of oxyntomodulin (19-37) (ED50 approximately 70 pmol/kg) was the same as that of oxyntomodulin (ED50 approximately 35 nmol/kg) after correction of the curve for the MCR (two-fold higher for the fragment). Although the dose-response curve for oxyntomodulin-(30-37) was not entirely parallel to that of the two other peptides, its maximal inhibition (at 15 nmol/kg) was the same as that of oxyntomodulin-(19-37). Thus, oxyntomodulin-(30-37) was an efficient as the longer molecules but approximately 150-fold less potent than oxyntomodulin. The MCR for oxyntomodulin-(30-37) was approximately 250 fold higher than that of oxyntomodulin, partly explaining the difference in potency. The (32-37) fragment, which is naturally released from the endocrine pancreas, was devoid of activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Miniglucagon [glucagon-(19-29)] is a component of the positive inotropic effect of glucagon.

Glucagon is well known for its cardiotonic effect, but its mechanism of action remains undetermined. In the present study, we showed that glucagon, under minimal degradation conditions, had no effect on the amplitude of contractility of beating chick embryo ventricular cells. This raised the question of the contribution of the active metabolite of glucagon, glucagon-(19-29), referred to as miniglucagon, to the positive inotropic effect of glucagon. Incubation of glucagon with heart cells led to its rapid conversion into miniglucagon, as measured by radioimmunoassay. Accumulation of the metabolite was maximal after 8 min and remained stable until 15 min. reaching 6% of the initial glucagon concentration. Bacitracin inhibited this processing of glucagon into miniglucagon. Miniglucagon, from 0.1 pM to 1 nM, exerted a potent negative inotropic action. The most striking observation was a 45% increase in the amplitude of cell contractility elicited by the combination of 30 nM glucagon with 1 nM miniglucagon. A similar effect was obtained when glucagon was replaced by a low concentration (75 microM) of 8-bromoadenosine 3',5'-cyclic monophosphate. We conclude that glucagon processing into miniglucagon may be essential for the positive inotropic effect of glucagon on heart contraction.

Animals↗
PubMed1990

Glucagon-(19-29), a Ca2+ pump inhibitory peptide, is processed from glucagon in the rat liver plasma membrane by a thiol endopeptidase.

Glucagon-(19-29) is 1000-fold more potent that glucagon as an inhibitor of the liver plasma membrane calcium pump, which suggests that this peptide fragment is naturally occurring. Since glucagon-(19-29) is undetectable in plasma, the processing of glucagon into its (19-29) fragment may occur upon interaction of glucagon with its target tissues. The use of a specific radioimmunoassay for glucagon-(19-29) in association with the separation and identification of peptides by high performance liquid chromatography revealed that, upon incubation at 37 degrees C with hepatic plasma membranes, glucagon is processed into its (19-29) C-terminal fragment. The identity of the fragment was confirmed by amino acid sequencing. The processing activity was inhibited by reagents of the thiol group and by 1,10-phenanthroline, suggesting that a thiol endopeptidase containing a catalytically active metal is involved in this processing. Following its production, glucagon-(19-29) was degraded with a half-life of less than 10 s. This degradation was inhibited by bacitracin and by the aminopeptidase inhibitors bestatin and amastatin. When glucagon was incubated with liver plasma membranes in the absence of inhibitors, the accumulation of glucagon-(19-29) reached a maximum at 2 min (1% of initial glucagon), followed by a slow decline. In the presence of bacitracin and bestatin, the amounts of glucagon-(19-29) obtained from glucagon increased continuously, 1 and 2% of glucagon being transformed after 10 and 30 min, respectively. The production of glucagon-(19-29) did not appear to be associated with the binding of glucagon to its receptors, since (i) guanosine 5'-(3-O-thio)triphosphate, a compound which decreases the glucagon-receptor interaction, could not decrease the conversion of glucagon into glucagon-(19-29); (ii) a glucagon analogue which displays a strongly decreased affinity for the hepatic glucagon receptors was processed similarly to glucagon. The conversion also occurs upon incubation with intact hepatoma cells in monolayer culture. These observations suggest that, under physiological conditions, glucagon is processed in liver by cleavage of the Arg17-Arg18 basic doublet, leading to the production of a fragment which is known to display an original biological specificity, namely the modulation of the hepatocyte plasma membrane calcium pump.

Adenosine Triphosphate↗
PubMed1990

Metabolic clearance rates of oxyntomodulin and glucagon in the rat: contribution of the kidney.

The half-life (t1/2) and metabolic clearance rate (MCR) of exogenous natural porcine oxyntomodulin (porcine OXM) and the synthetic analog of rat oxyntomodulin, [Nle27]-OXM (rat OXM), were compared with that of glucagon in control, sham-operated and acutely nephrectomized rats using the primed-continuous infusion technique. The half-disappearance times for porcine OXM (8.2 +/- 0.5 min) and rat OXM (6.4 +/- 0.5 min) were 3-fold slower than that of glucagon (1.9 +/- 0.1 min). Acute bilateral nephrectomy significantly prolonged the half-disappearance time of rat OXM (8.2 +/- 0.7 min) and glucagon (3.6 +/- 0.4 min) compared with that of sham-operated animals (6.5 +/- 0.8 min and 2.5 +/- 0.2 min, respectively). The mean MCRs were similar for porcine and rat OXM (11.3 +/- 0.7 and 11.9 +/- 0.5 ml.kg-1.min-1) but were 3 times lower than that measured with glucagon (36 +/- 5 ml.kg-1.min-1). Bilateral nephrectomy reduced the MCR of OXM and glucagon by 38% and 34%, respectively. No significant increase in C-terminal glucagon immunoreactivity was noticed during infusion of either porcine or rat OXM, measured directly in plasma, with a specific C-terminal glucagon antiserum or after HPLC. In the course of the glucagon infusion, blood glucose was increased 2-fold, while the same dose of porcine OXM or of rat OXM induced only a small increase over the values in phosphate buffer-infused rats. 10 times higher doses of rat OXM were necessary to obtain a similar hyperglycemic effect. These results indicate that: (1) the metabolism of OXM is 3-fold slower than that of glucagon, (2) renal clearance contributed close to 35% of the overall metabolic plasma extraction for OXM and glucagon and (3) OXM, although effective at a higher dose, when compared with glucagon, displays a hyperglycemic effect probably through the glucagon receptors.

Animals↗
PubMed1990

Glucagon-(19-29) exerts a biphasic action on the liver plasma membrane Ca2+ pump which is mediated by G proteins.

We have recently shown that nanomolar concentrations of glucagon-(19-29), which can derive from native glucagon by proteolytic cleavage of the dibasic doublet Arg17-Arg18, inhibit the Ca2+ pump in liver plasma membrane vesicles independently of adenylyl cyclase activation (Mallat, A., Pavoine, C., Dufour, M., Lotersztajn, S., Bataille, D., and Pecker, F. (1987) Nature 325, 620-622). We report here that the regulation of the Ca2+ pump by glucagon-(19-29) is dependent on guanine nucleotides. In the presence of 10 microM guanosine 5'-3-O-(thio) triphosphate (GTP gamma S) or 75 microM GTP, glucagon-(19-29) caused a biphasic regulation of the Ca2+ pump. ATP-dependent Ca2+ transport was inhibited in the presence of 10 pM to 1 nM glucagon-(19-29), while higher concentrations of the peptide (1-100 nM) reversed the inhibition caused by lower ones. GTP gamma S alone, at high concentrations (100 microM), reproduced the inhibitory effect of glucagon-(19-29) and induced a 40% inhibition of the basal activity of the Ca2+ pump which was reversed by low concentrations of glucagon-(19-29) (10 pM to 1 nM). Treatment of rats with cholera toxin resulted in a 70% increase in the basal activity of the Ca2+ pump, a loss of sensitivity to GTP gamma S and to the biphasic regulation by glucagon-(19-29). Treatment with pertussis toxin did not affect the response of the Ca2+ pump to GTP gamma S and glucagon-(19-29). We conclude that glucagon-(19-29) can exert a biphasic effect on the Ca2+ pump which is mediated by G protein(s) sensitive to cholera toxin.

Adenylate Cyclase Toxin↗
PubMed1990

[Post-translational maturation of peptides of the glucagon family. Relationship with their mode of action].

Two types of proglucagon processing have been evidenced in producing tissues (endocrine pancreas, stomach, intestine, central nervous system) using antibodies recognizing the epitopes unmasked during processing, which takes place at dibasic sites. A first type, leading essentially to glucagon, has been observed in the two former tissues; a second type, leading to peptides (oxyntomodulin and glicentin) containing an additional C-terminal octapeptide, has been shown in the two latter. All peptides are released in plasma and reach their targets: liver, fat... (control of metabolism) for glucagon and gastric mucosa (control of acid secretion) for the octapeptide-bearing peptides. The mode of action of these peptides includes receptors coupled to adenylate cyclase and a processing, at a dibasic site, of the circulating peptides leading to C-terminal fragments which act through non cyclic AMP-dependent mechanisms, such as the control of the plasma membrane calcium pump.

Animals↗
PubMed1989

Membrane-cytoskeleton dynamics in rat parietal cells: mobilization of actin and spectrin upon stimulation of gastric acid secretion.

The gastric parietal (oxyntic) cell is presented as a model for studying the dynamic assembly of the skeletal infrastructure of cell membranes. A monoclonal antibody directed to a 95-kD antigen of acid-secreting membranes of rat parietal cells was characterized as a tracer of the membrane movement occurring under physiological stimuli. The membrane rearrangement was followed by immunocytochemistry both at the light and electron microscopic level on semithin and thin frozen sections from resting and stimulated rat gastric mucosa. Double labeling experiments demonstrated that a specific and massive mobilization of actin, and to a lesser extent of spectrin (fodrin), was involved in this process. In the resting state, actin and spectrin were mostly localized beneath the membranes of all cells of the gastric gland, whereas the bulk of acid-secreting membranes appeared diffusely distributed in the cytoplasmic space of parietal cells without any apparent connection with cytoskeletal proteins. In stimulated cells, both acid-secreting material and actin (or spectrin) extensively colocalized at the secretory apical surface of parietal cells, reflecting that acid-secreting membranes were now exposed at the lumen of the secretory canaliculus and that this insertion was stabilized by cortical proteins. The data are compatible with a model depicting the membrane movement occurring in parietal cells as an apically oriented insertion of activated secretory membranes from an intracellular storage pool. The observed redistribution of actin and spectrin argues for a direct control by gastric acid secretagogues of the dynamic equilibrium existing between nonassembled (or preassembled) and assembled forms of cytoskeletal proteins.

Actins↗
PubMed1989

1H n.m.r. conformational studies on the C-terminal octapeptide of oxyntomodulin, a beta-turn locked by a salt bridge.

The octapeptide Lys-Arg-Asn-Lys-Asn-Asn-Ile-Ala (Arg4 in the human sequence) is the C-terminal part of porcine oxyntomodulin, an endogeneous peptide which is a potent inhibitor of stimulated acid secretion. This octapeptide exhibits the whole range of biological activities of the parent hormone. In the present work we report an 1H n.m.r. investigation of the conformational properties of the octapeptides of pig and human sequences in dimethylsulfoxide-d6 (DMSO) solution. The various resonances were assigned on the basis of two-dimensional COSY and NOESY experiments. Other experiments such as (i) temperature and concentration dependence of the amide proton chemical shifts, (ii) effects of ionic strength, (iii) comparison of the spectra with different analogues, were performed. We showed that in DMSO, the conformation of the octapeptide is directly related to the ionisation state of the C-terminus carboxyl group of alanine. In carboxylic state, the peptide adopts an extended conformation, while in the carboxylate state the four last residues (Asn-Asn-Ile-Ala) are involved in a type II beta-turn structure probably locked by a salt bridge between the carboxyl group of Ala8 and the epsilon ammonium group of Lys4 (or the guanidinium group of Arg4). These observations provide an insight into the possible conformational tendencies of this peptide in biological media.

Amino Acid Sequence↗
PubMed1989

Inhibitory effect of the C-terminal octapeptide of oxyntomodulin on pentagastrin-stimulated gastric acid secretion in man.

The effect of the C-terminal octapeptide of human oxyntomodulin (KA-8) on pentagastrin-stimulated gastric acid secretion was studied versus placebo in eight human volunteers after an overnight fast, using a double-blind randomized protocol. KA-8 (7.5 nmol/kg body weight) was bolus-injected at a plateau stimulation obtained with pentagastrin (3 micrograms/kg/h). KA-8 significantly decreased the volume secreted by 36% (p less than 0.025) and the acid output by 32% (p less than 0.025) 20 min after injection. The areas under the curve were significantly reduced for volume and acid output during the 1st h after peptide injection.

Adult↗