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J C Meunier

Publications and source records attributed to J C Meunier.

At least 73 records · Page 4Linked to original sources
PubMed1992

Increased calcitonin gene-related peptide- and cholecystokinin-like immunoreactivities in spinal motoneurones after dorsal rhizotomy.

Possible changes in neuropeptides within the ventral horn of the spinal cord were investigated after unilateral dorsal rhizotomy at the lumbar level (L1-L6) in adult rats. Ten days after the surgery, immunohistochemical observations and radioimmunological determinations confirmed a marked loss of calcitonin gene-related peptide (CGRP)- and substance P (SP)-like immunoreactivities within the superficial layers of the deafferented dorsal horn, as expected from the degeneration of primary afferent fibres containing these peptides. A concomitant increase in immunohistochemical staining and levels of CGRP (+296%) and CCK (+71%)-like immunoreactivities was observed in the ipsilateral ventral horn where both peptides are located in motoneurones. In contrast, substance P-like immunoreactivity that is confined to fibres and terminals within the ventral horn, was not altered by dorsal rhizotomy. These data indicate that the expression of neuropeptides in spinal motoneurones can be influenced by primary afferent inputs.

Animals↗
PubMed1992

An improved method for the purification of lignin peroxidases from Phanerochaete chrysosporium INA-12: properties of two major isoforms.

1. Phanerochaete chrysosporium INA-12 secretes several lignin peroxidase isoenzymes. This paper reports an improved procedure for the purification of the different isoforms compared to those previously described. 2. Lignin peroxidases are first concentrated and prefractionated on fast-flow ion-exchangers which avoid concentration by ultrafiltration and dialysis. 3. Further purification is achieved by hydrophobic interaction chromatography and anion-exchange FPLC. 4. Two major forms were purified to homogeneity. Kinetic measurements and protein characterization (isoelectric points, phosphate content) suggest that they are similar to those produced by P. chrysosporium BKM strain.

Basidiomycota↗
PubMed1992

Purification and properties of factor XIII from human placenta.

1. FXIII was isolated and purified over 4000 fold from human placenta to apparent electrophoretic homogeneity by a new procedure including ethanol precipitation. DEAE-Cellulose, molecular sieving on Sephacryl S-300 and Phenyl-Sepharose chromatography. 2. Its pI was about 5.1. Under appropriate conditions, the incubation of FXIII in the presence of thrombin did not lead to inactivation cut in the polypeptidic chain. 3. FXIII was also activated by CaCl2 and, in a lesser extent, by other divalent cations like SrCl2, BaCl2 or MgCl2. 4. The binding of calcium to FXIII exhibited a negative cooperativity. 5. The activity-pH curve of the calcium-activated enzyme did not appear very different from that of the thrombin-activated enzyme.

Calcium↗
PubMed1992

Effect of temperature and pH on factor XIIIa from human placenta.

1. The activation of the native enzyme was achieved by a proteolytic procedure involving thrombin. 2. The pH profile was independent of the nature of the substrates assayed (casein or dimethylcasein plus putrescine). The optimum pH was between 7.6 and 7.9 and the pK values were 6.5/7 and 8.7/9. A cysteinyl residue appeared to be involved in the pH-dependence activity. 3. In the presence of calcium, the thermostability of enzyme was increased: the temperature at which enzyme lost half of its activity increased up to 7 degrees C. 4. The kinetics of the thermal deactivation of F XIIIa depended on the presence or absence of calcium. 5. In its presence the reaction obeyed second order kinetics, while in its absence, the kinetics were of first order. In the first case, the irreversible thermal deactivation could be described by a two-step mechanism (N----X----D) while in the second case, the deactivation followed the simple model (N----D). 6. Neither divalent cations like Sr2+, Ba2+, Mg2+, nor bovine serum-albumin and polyhydric alcohols were able to increase the thermostability of F XIIIa. 7. Thermal deactivation of F XIIIa did not appear linked to the redox state of enzyme, nor to the modification of SH groups. 8. We observed a good correlation between the loss of activity and the unfolding of the polypeptide chain of F XIIIa during heating. 9. The optimum temperature of F XIIIa activity was 40 degrees C at pH 8 and 45 degrees C at pH 7.

Calcium↗
PubMed1992

Two-sites immunoradiometric assay using monoclonal antibodies for the determination of serum human sex hormone binding globulin.

An immunoradiometric assay (IRMA) for the determination of human Sex Hormone Binding Globulin (SHBG) in serum is described. This IRMA uses three mouse monoclonal antibodies. Two monoclonals anti-human SHBG are coated on tubes and used as capture antibodies. The third monoclonal labeled with 125I completes the system, allowing the formation of the "sandwich". The detection limit of the assay is 2.5 femtomol SHBG per tube (250 pg/tube). Using this test for the measurement of SHBG and radioimmunoassays for the determination of total Testosterone and Estradiol, we calculated the Free Androgen Index (FAI) and the Free Testosterone. The results obtained were compared with the values of Free Testosterone measured by equilibrium dialysis. There is a close correlation between both calculated parameters and the levels of Free Testosterone, validating this SHBG assay.

Antibodies, Monoclonal↗
PubMed1992

[Opioid receptors, tolerance and dependence].

Opioid substances act, both in the central and peripheral nervous systems, by regulating--via a specific receptor of the mu, delta or kappa type and a G protein--the activity of either of three recognized cellular effectors: adenylate cyclase, a K+ channel and a Ca++ channel. In the short term, opioid effects are of a neuromodulatory origin: they are likely to reflect opioid inhibition of neurotransmitter release (e.g.: of substance P in the spinal cord). In the long term, opioids induce the two adaptative phenomena known as tolerance and dependence whose mechanisms are considerably less well understood. Tolerance refers to a decreased responsiveness to the opioid upon repeated administration of the drug. Tolerance might reflect desensitization, a process which involves uncoupling of opioid receptor and of G protein (in the case of homologous desensitization) and, possibly, down-regulation of a G protein which the opioid receptor shares with other types of receptor (heterologous desensitization). Dependence refers to a latent, opioid-induced physiological state which expresses itself as a typical excitation syndrome upon withdrawal of the drug or administration of an opioid antagonist. The molecular events whereby dependence develops are unknown, yet they may involve (i) adenylate cyclase supersensitivity, an adaptation reaction where upon withdrawal of the inhibitory opioid induces an enhanced activity of the enzyme and, (ii) increased release of neurotransmitter (presynaptic facilitation).

Down-Regulation↗
PubMed1991

Properties of oxidized and reduced spinach (Spinacia oleracea) chloroplast fructose-1,6-bisphosphatase activated by various agents.

Fructose-1,6-bisphosphatase (FBPase) can be reduced and activated by either dithiothreitol or reduced thioredoxin. This activation is pH-dependent. An amino acid group with a pK value of 5.55 is involved in this process. Both enzyme forms can also be stimulated by agents such as fructose 1,6-bisphosphate, Mg2+, Ca2+ and Ca2+/fructose 1,6-bisphosphate. FBPase reduced by dithiothreitol is more strongly activated than the enzyme reduced by thioredoxin. The specificity constant (kcat./Km) is enhanced over 2.5-25-fold and 1.5-2-fold (depending on the agent used) for FBPase reduced by dithiothreitol and thioredoxin respectively. In both cases, no new kinetic properties appeared. The pH-activity profile of the stimulated enzyme is slightly shifted towards the acidic side with respect to the reduced enzyme. A lag phase is observed in the progress curve of both enzymic forms, treated or untreated. Each agent used to stimulate must induce a new conformation of the enzyme, more active than the initial one, characterized by a specificity constant and a relaxation time. This lag phase tends to disappear when the assay temperature is increased. Temperature has the same effect on the activity of oxidized, reduced and stimulated FBPase, but different effects on the stability of the different forms.

Calcium↗
PubMed1991

Non reductive activation of spinach chloroplastic fructose-1,6-bisphosphatase: evidence for structural modification of the enzyme.

Preincubation of chloroplastic fructose-1,6-bisphosphatase (FBPase) in the presence of Ca2+/fructose-1,6-bisphosphate (FBS) gives rise to an active enzyme. This non-reductive activation at pH 8 occurs in the same range of time (min) as the well known reductive activation by thioredoxins and this process is reversible. A conformational change of the enzyme occurs upon the activation by Ca2+/FBP. Indeed, the circular dichroism and the fluorescence spectra of the inactive and active enzymes are different. The titration of the sulfhydryl groups of both enzymes indicates that one -SH group per monomer is unmasked upon activation, and the isoelectrofocusing pattern shows that the pI of inactive FBPase is shifted from 4.26 to 4.56 upon this non-reductive process.

Calcium↗
PubMed1990

Regulation by Na+ ions and GppNHp of the interaction between a G protein and the amphibian type of opioid receptor.

Digitonin treatment of frog brain membranes in 50 mM Tris-HCl yields a soluble extract that contains nearly equal amounts of free and G protein-bound opioid receptor molecules (Mollereau et al., 1988, J. Biol. Chem. 263, 18003). We report here that the balance of the two forms of the opioid receptor in digitonin solution is dependent on the environment of the membrane suspension at the time of solubilization with the detergent. Preincubating the membrane suspension with 50 microM GppNHp or with 120 mM NaCl results, in the two cases, in a digitonin extract that no longer displays the G protein-bound form of the receptor, i.e., the form of the receptor which exhibits high affinity for the opiate agonist etorphine in binding studies, as well as large apparent molecular size in sucrose gradients. Assuming that the situation in soluble extracts faithfully reflects the one in the membrane, these results would exclude the possibility that in a physiological environment the opioid receptor is in part precoupled with a G protein in the absence of an opioid agonist.

Animals↗
PubMed1990

Apparent precoupling of kappa- but not mu-opioid receptors with a G protein in the absence of agonist.

Rabbit and guinea-pig cerebellum membranes contain a very high (greater than 80%) proportion of mu- and kappa-opioid receptors, respectively. Rabbit (mu) and guinea-pig (kappa) cerebellum membranes were (i) labeled either with the opiate agonist, [3H]etorphine (Kd = 0.1-0.2 nM), or with the opiate antagonist, [3H]diprenorphine (Kd = 0.1 nM), in the absence or presence of Na+ and/or 5'-guanylylimidodiphosphate (GppNHp), (ii) solubilized with digitonin (1%, w:v) and (iii) the radioactivity in the soluble extracts analyzed by ultracentrifugation in sucrose gradients. In the soluble extracts from rabbit cerebellum (mu) membranes, bound [3H]etorphine sedimented faster (S20,w congruent to 12S) than bound [3H]diprenorphine (10S), while in those from guinea-pig cerebellum (kappa) membranes, bound [3H]etorphine and bound [3H]diprenorphine sedimented at the same position (12S). Na+ selectively decreased recovery of the bound tritiated agonist in the two soluble preparations. When they had been generated in the presence of GppNHp but in the absence of Na+, the [3H]etorphine complexes of the mu- and kappa-opioid receptors as well as the [3H]diprenorphine complex of the kappa-opioid receptor were all recovered at position 10S, indicating that GppNHp had induced a decrease of the apparent molecular size of the two types of opioid receptors. These data are interpreted in terms of mu- and kappa-opioid receptors being capable of physically interacting with a G protein (GTP binding regulatory protein) yet, unlike the mu-opioid receptor which does so only in the presence of an agonist, the kappa-opioid receptor appears to be precoupled with a G protein.

Animals↗
PubMed1990

Molecular mechanisms of the irreversible thermal denaturation of guinea-pig liver transglutaminase.

When transglutaminase is heated at temperatures above 40 degrees C, it loses its activity according to a two-step mechanism [Nury, Meunier & Mouranche (1989) Eur. J. Biochem. 180, 161-166]: N----X(TD)----D However, the nature of the molecular events responsible for the irreversible denaturation is still unknown. Investigation of the effects of dithiothreitol and 5,5'-dithiobis-2-nitrobenzoate on the kinetics of inactivation, titrations of ammonia released by deamidation and of thiol groups on the native and denatured enzymes and SDS/PAGE rule out the involvement of covalent processes during the denaturation of transglutaminase at 55 degrees C and pH 7. Of the two possible kinds of non-covalent events, i.e. unfolding of the polypeptide chain and aggregation of enzyme molecules, we show that both occur, though only the former process is responsible for the denaturation. The latter process, aggregation, follows the unfolding of the molecules.

Animals↗
PubMed1990

Morphine metabolism in the naturally morphine-tolerant afghan pika: a preliminary study.

The afghan pika (Ochotona rufescens), a lagomorph which is naturally tolerant to the analgesic action of morphine, metabolizes morphine into morphine 3-glucuronide apparently faster than does the rabbit, another lagomorph which is however normally responsive to morphine. In the two species, following morphine administration, another unidentified component appears very soon (5 min) in pika blood plasma and much later (60 min) in rabbit blood plasma. This unknown component which appears not to be morphine derived might be involved in the natural resistance of the Afghan pika to morphine.

Animals↗
PubMed1990

Effects of calcium on the stability and activity of guinea pig liver transglutaminase.

The binding of calcium to transglutaminase was studied by a kinetic method and by spectrophotometric titration. By the first method, we have shown that the binding of a single calcium per molecule, with a binding constant of 7500 +/- 1300 M-1 (at 55 degrees C), was responsible for the enhancement of the thermostability. The kinetic constants of the deactivation for the unliganded native form and the liganded native one are 1.47 +/- 0.04 min-1 and 0.32 +/- 0.05 min-1 respectively. The enhancement of thermostability is due to the stabilization of the native form, since the deactivation constants of the liganded and unliganded intermediate forms are equal. The total number of calcium binding sites, determined by titration is 4, and therefore, only 1 of them should be implicated in the thermostability. The 4 apparent association constants have been determined by a non-linear fitting of the data to the Adair equation. We have also shown a positive co-operative behaviour of the transglutaminase when the transferase is monitored versus calcium concentration.

Animals↗
PubMed1990

Photoactivatable opiate derivatives as irreversible probes of the mu-opioid receptor.

The synthesis of aryldiazonium and arylazido derivatives of carfentanil, etonitazene, and naltrexone and of a triazaspirodecane derivative is described. The chemical stability and the spectral characteristics of these compounds were verified, and their binding affinity constants for the different opioid receptor classes were determined, in the absence of light, from competition experiments. With the exception of the naltrexyl derivatives, which remained nonselective, all compounds tested displayed a pronounced mu-binding selectivity with mu/delta and mu/kappa ratios ranging from 12 to 1000. After irradiation, only the arylazido probes led to an irreversible mu-binding-site inactivation. This inactivation fulfilled the criteria for photoaffinity labeling such as protection against inactivation by other opiate ligands and absence of an effect of scavengers on the extent of the inactivation. Most of the photoactivatable probes formed long-lasting reversible complexes with the opioid binding sites: an efficient dissociation procedure was thus required to discriminate between pseudoirreversible and covalent complexes. The marked differences in labeling efficacy between aryldiazonium salts and their corresponding arylazido derivatives are discussed.

Affinity Labels↗
PubMed1990

mu-Opioid receptors and not kappa-opioid receptors are coupled to the adenylate cyclase in the cerebellum.

The putative regulatory effect of opioids on adenylate cyclase was investigated in two different preparations containing, respectively, two different populations of opioid receptors: the rabbit cerebellum (greater than 75% mu-opioid receptors) and the guinea pig cerebellum (greater than 80% kappa-opioid receptors). In the mu-preparation, but not in the kappa-preparation, opioids inhibited the basal and the forskolin-stimulated adenylate cyclase activity in a dose-dependent manner and stereospecifically. The inhibition was in the 20-30% range, required the presence in the assay medium of Mg2+ and of GTP, but was independent of the presence of Na+. Pharmacological characterization of the inhibitory response in the rabbit cerebellum clearly showed that it was under the control of a mu-opioid binding site, with the effect being elicited by non-selective (etorphine and morphine) and mu-selective (Tyr-D-Ala-Gly-Me-Phe-Gly-ol) agonists, whereas delta- and kappa-selective agonists were almost totally ineffective. ADP ribosylation of inhibitory GTP-binding protein by pertussis toxin failed to block the inhibitory effect of opioids, and data presented suggest that this failure is likely to be the consequence of a limited access of the toxin to its substrate in rabbit cerebellum membranes.

Adenylate Cyclase Toxin↗
PubMed1989

Kinetic study of the irreversible thermal denaturation of Bacillus licheniformis alpha-amylase.

The irreversible thermal inactivation of Bacillus licheniformis alpha-amylase was studied. A two-step behaviour in the irreversible denaturation process was found. Our experimental results are consistent only with the two-step model and rule out the two-isoenzyme one. They suggest that the deactivation mechanism involves the existence of a temperature-dependent intermediate form. Therefore the enzyme could exist in a great number of active conformational states. We have shown that Ca2+ is necessary for the structural integrity of alpha-amylase. Indeed, dialysis against chelating agents leads to a reversible enzyme inactivation, though molecular sieving has no effect. Further, the key role of Ca2+ in the alpha-amylase thermostability is reported. The stabilizing effect of Ca2+ is reflected by the decrease of the denaturation constants of both the native and the intermediate forms. Below 75 degrees C, in the presence of 5 mM-CaCl2, alpha-amylase is completely thermostable. Neither other metal ions nor substrate have a positive effect on enzyme thermostability. The effect of temperature on the native enzyme and on one intermediate form was studied. Both forms exhibit the same optimum temperature. Identical activation parameters for the hydrolytic reaction catalysed by these two forms were found.

Bacillus↗
PubMed1989

Binding of dynorphin A and related peptides to kappa- and mu-opioid receptors: sensitivity to Na+ ions and Gpp(NH)p.

We have examined the effects of Na+ ions and 5'-guanylyl imidodiphosphate (Gpp(NH)p) on the equilibrium binding of dynorphin A and of a series of related agonist and antagonist peptides to kappa- and mu-opioid receptors in guinea pig (kappa) and rabbit (mu) cerebellum membrane preparations. The binding to kappa sites of dynorphin A and of the peptides displaying agonist properties was strongly inhibited in the presence of 120 mM NaCl and 50 microM Gpp(NH)p. In contrast, a somewhat lower sensitivity to the inhibitory effect of the two allosteric effectors was observed for the analogues of the series showing antagonist properties. The same general behavior, but more marked, was observed at mu sites, at both mu- and kappa-opioid receptors. The peptides had biochemical properties (binding sensitivity vs. insensitivity to sodium ions and guanine nucleotides) that correlated well with their biological activities (agonist vs. antagonist) previously determined in in vitro pharmacological bioassays.

Animals↗