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M Potier

Publications and source records attributed to M Potier.

At least 127 records · Page 7Linked to original sources
PubMed1986

Solubilization and characterization of D2-dopamine receptors in an estrone-induced, prolactin-secreting rat pituitary adenoma.

D2-dopamine (3,4-dihydroxyphenylethylamine) receptors were successfully solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate from an estrone-induced rat pituitary adenoma. Forty-five percent of initial protein and 48% of initial [3H]spiroperidol binding sites were solubilized. The high affinity as well as the stereoselectivity of the sites was preserved. The order of potency of dopaminergic agonists was found to be typical of D2 receptors. Target size analysis by radiation inactivation indicated a molecular weight of 143,000 +/- 3,000 and of 106,000 +/- 4,000 daltons for membrane-bound and solubilized receptors, respectively. This suggests the loss of a 37,000-dalton subunit during solubilization without significant modification of binding characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of receptor protein preparation photolabeled with N-(p-azido-m[125I]iodophenethyl)spiroperidol confirmed the existence of a 94,000-dalton peptide which probably constitutes the ligand binding site of the receptor. Thus, our data indicate that chronic estrogen treatment of rats, although inducing a pituitary adenoma, does not modify the pharmacological characteristics of D2 receptors. These data suggest therefore that these adenoma may represent an ideal source of material for further biochemical characterization of D2 receptors.

Adenoma↗
PubMed1986

Characterization of the mutant N-acetylglucosaminylphosphotransferase in I-cell disease and pseudo-Hurler polydystrophy: complementation analysis and kinetic studies.

Complementation was examined among various types of I-cell disease and pseudo-Hurler polydystrophy by monitoring N-acetylglucosaminylphosphotransferase activity in multinucleated cells produced by fusing pair combinations of cultured skin fibroblasts. Patients with the classical forms of these disorders (5 I-cell disease and 3 pseudo-Hurler polydystrophy cell lines) comprised one complementation group and 5 cell lines from patients with variant forms of pseudo-Hurler polydystrophy comprised a distinct complementation group. In the first group, total or partial deficiency of the transferase activity was demonstrated with both natural (lysosomal enzymes) and artificial (alpha-methylmannoside) acceptor substrates with low Vmax but apparently normal Km values for the donor (UDP-GlcNAc) and acceptor (alpha-methylmannoside) substrates. The activity toward artificial substrate could be inhibited by adding exogenous lysosomal enzyme preparations to the reaction mixture. In the second group, the cells demonstrated deficiency of the transferase activity toward lysosomal enzyme acceptors but had normal activity toward alpha-methylmannoside acceptor and this activity could not be inhibited by the addition of exogenous lysosomal enzyme preparations. These findings suggest that N-acetylglucosaminylphosphotransferase is composed of at least two distinct subunits, a catalytic subunit which is absent or defective in the first complementation group, and a recognition subunit which is altered or deficient in the second group.

Cells, Cultured↗
PubMed1985

Cholesteryl ester transfer protein. Size of the functional unit determined by radiation inactivation.

Radiation inactivation was used to determine the functional Mr of cholesteryl ester transfer protein (CETP) in rabbit plasma from control and irradiated animals. This technique reveals the size of the functional unit required to carry out the transfer function. The functional Mr was calculated to be 70 000 +/- 3000 (mean +/- SD) for both control and irradiated rabbits. This result is in accordance with the Mr obtained by a completely different method, namely SDS-polyacrylamide gel electrophoresis of a partially purified (110-fold) rabbit CETP. The pI of this CETP was found by isoelectric focusing to be equal to 5.95. The results suggest that the functional unit of this enzyme is the monomer.

Animals↗
PubMed1985

Regulatory proteins (inhibitors or activators) affect estimates of Mr of enzymes and receptors by radiation inactivation. A theoretical model.

The radiation-inactivation method allows the determination of the Mr of enzymes and receptors by monitoring the decay of biological activity as a function of absorbed dose. The presence of regulatory or effector proteins (inhibitors or activators) associated with an enzyme or receptor, or released in the preparation after tissue homogenization, may affect the decay of biological activity. How the activity is affected, however, will depend on the type of inhibition (competitive or non-competitive), the inhibitor or activator concentration, the dissociation constant of the enzyme-effector system, and the effector Mr relative to that of the enzyme. Since little is known on how effector proteins influence radiation inactivation of enzymes and receptors, we have considered a theoretical model in an effort to provide a framework for the interpretation of experimentally obtained data. Our model predicts that competitive and non-competitive inhibitors of enzymes could be distinguished by analysing irradiated samples with various substrate concentrations. Inhibitors will decrease whereas activators will increase the apparent target size of enzymes or receptors.

Binding, Competitive↗
PubMed1985

Temperature dependence of the radiation inactivation of proteins.

The radiation inactivation method allows determination of the relative molecular mass (Mr) of proteins by exposure to high doses of ionizing radiation. The analysis by target theory of biological activity decay curves yields the size of the protein. A correction factor for Mr has been routinely used in the literature when irradiation is conducted at low temperature. Since the radiation inactivation of proteins is affected by temperature, we propose a general equation which relates Mr of a protein to D37,t, the dose in megarads at a given temperature t (in degree C) where 37% of its initial biological activity remains log Mr = 5.89 - log D37,t - 0.0028t. It is concluded that temperature affects the amount of absorbed radiation energy required to inactivate 1 mol of protein.

Mathematics↗
PubMed1985

Oral and dental development in X chromosome aneuploidy.

Though frequently noted, orofacial anomalies in poly X syndromes have been seldom reviewed. In a study of patients with an X chromosome aneuploidy (XXXXX, XXXX, XXXXY, XXXY) the following facial and dental defects were found to be characteristic features of these syndromes: mainly bifid uvula and macroglossia, enamel defects, dental taurodontism and abnormal roots. These observations are discussed in the light of the possible role of X-linked genes.

Adolescent↗
PubMed1985

Molecular weight of human brain neutral sphingomyelinase determined in situ by the radiation inactivation method.

The radiation inactivation method was used to determine the molecular weight of membrane-bound neutral sphingomyelinase from normal human brain. Inactivation curves showed a molecular mass of 167,000 +/- 32,000. Molecular weights of two control enzymes, beta-N-acetylglucosaminidase and nonspecific beta-glucosidase, determined by the same procedure, were consistent with previous reports.

Acetylglucosaminidase↗
PubMed1984

Radiation inactivation of membrane proteins: molecular weight estimates in situ and after Triton X-100 solubilization.

Target size analysis by radiation inactivation is widely used for molecular weight determination of membrane enzymes and receptors in situ without the need for prior solubilization or purification. However, since most molecular weight data available in the literature on membrane proteins involve the use of detergents for solubilization, the target sizes of membrane proteins in situ and after solubilization by detergent treatment have been compared. Using data from the literature and personal results, three different types of behavior of membrane proteins in presence of detergents were found: (i) uncoupling of subunits (electric eel acetylcholinesterase, placental steroid sulfatase, and human nonspecific beta-glucosidase); (ii) coupling of protein molecules (mouse liver neuraminidase, and rat liver insulin receptor regulatory component); and (iii) no major change in quaternary structure (rat liver insulin receptor, kidney gamma-glutamyltransferase, asialoglycoprotein receptor, insulin degrading enzyme, and human leucocyte neuraminidase). For all these proteins, there is a statistically significant increase in target size of about 24% over the value obtained in situ without detergent. A relatively large body of literature data involving a variety of membrane proteins, membrane types, and irradiation conditions (electron accelerators or 60Co sources, and proteins irradiated in lyophilized form or frozen solution) was examined, and it was concluded that target sizes of membrane proteins, irradiated in the presence of Triton X-100, should be diminished by a factor of about 24% to obtain the molecular weight value.

Animals↗
PubMed1984

Modifications of the molecular weight of membrane-bound nonspecific beta-glucosidase in type 1 Gaucher disease determined in situ by the radiation inactivation method.

The radiation inactivation method has been used to compare the molecular weight of the nonspecific membrane-bound beta-glucosidase in situ in normal human spleen and in that of two patients with Gaucher disease type 1. We report, in type 1 Gaucher spleen, the presence of a high molecular weight component (557 000) in addition to the normal low molecular weight component (97 800). The various possible hypotheses explaining this high molecular weight component are discussed.

Adult↗
PubMed1984

Friedreich's ataxia: malic enzyme activity in cellular fractions of cultured skin fibroblasts.

We have measured the activity of malic enzyme NADP+ dependent in the nuclear, mitochondrial, lysosomal and cytosolic fractions of cultured skin fibroblasts from twelve patients with Friedreich's ataxia and nine control subjects. Hexosaminidase, cytochrome-C-oxidase, lactate dehydrogenase and malic enzyme NAD+ dependent were used as marker enzymes. The activity of malic enzyme NADP+ dependent was not significantly reduced in the mitochondrial fraction of patients with Friedreich's ataxia as compared with controls. When corrected for possible contamination between mitochondrial and cytosolic fractions, malic enzyme NADP+ dependent activity was still not significantly reduced in patients with Friedreich's ataxia. Unless critical methodological differences were overlooked in this or previously published studies, we conclude that mitochondrial malic enzyme deficiency is not the primary genetic defect underlying Friedreich's ataxia.

Adolescent↗
PubMed1984

Date of conception and prevention of neural tube defects.

The authors studied the time of conception of 280 fetuses and liveborns with anencephaly or spina bifida seen in a French Canadian population during a period of six years. There is an evident seasonal variation in the conception of NTD fetuses. Using a wave statistical model applied to data recorded over a six year period the authors have rejected the "no variation between the six two-month intervals" hypothesis at a P less than or equal to 0.005 level. It is postulated that, in any vitamin intake or environmental program to prevent NTD in families at risk, one must take into account the annual and seasonal variations, ethnic groups identity and the date of conception in the choice of control mothers.

Canada↗
PubMed1984

Differential stabilities of fetal intestinal disaccharidases determine their relative amounts released into amniotic fluid.

Comparison of relative disaccharidase activities in jejunal mucosa, amniotic fluid, and meconium samples taken from the ileum, proximal and distal colon of 5 human fetuses, indicates that trehalase and lactase are more sensitive to the denaturation conditions existing in the fetal intestinal lumen than maltase, sucrase, palatinase and turanase . Relative activities of sensitive disaccharidases (trehalase and lactase) were also low in the amniotic fluid of these fetuses as compared to the jejunal mucosa. These results suggest that the in vivo stability of disaccharidases, exposed to proteolytic digestion and other denaturing conditions in the fetal intestinal lumen, governs the relative activities of these enzymes released into the amniotic fluid.

Amniotic Fluid↗
PubMed1983

The molecular weight of the calcium-transport-ATPase of the human red blood cell determined by radiation inactivation.

Radiation inactivation was applied to analyze the molecular weight of the functional unit of (Ca2+ + Mg2+)-ATPase in human erythrocyte membranes. The enzyme activity was stable for at least 7 days at room temperature in membranes lyophilized in the presence of sucrose (150-300 mM). The enzyme activity in the lyophilized membranes and remaining after irradiation from a 60Co source was activated by calmodulin. A Mr of 290,000 +/- 15,000 was determined for (Ca2+ + Mg2+)-ATPase activity. Since the Mr by SDS-polyacrylamide gel electrophoresis is approximately 138,000 (Niggli et al. (1979) J. Biol. Chem. 254, 9955-9958), our results suggest that the Ca2+ pump ATPase functions as a dimer in the native human erythrocyte membrane.

Ca(2+) Mg(2+)-ATPase↗
PubMed1983

Modification of subunit interaction in membrane-bound acid beta-glucosidase from Gaucher disease.

The radiation inactivation method has been used to determine the molecular mass of membrane-bound acid beta-glucosidase (EC 3.2.1.21) in situ, in normal human spleen and in that of two patients with type I Gaucher disease: the molecular mass in Gaucher spleen is about double (125 000 +/- 8900) of that found in the normal spleen (67 000 +/- 7700) which is compatible with the existence of subunit coupling in the muted acid beta-glucosidase. From the results, we conclude that subunit interaction is altered in mutant acid beta-glucosidase and that this may be due to a direct effect of the mutation.

Cell Membrane↗
PubMed1983

Target size analysis by radiation inactivation: a large capacity tube rack for irradiation in a Gammacell 220.

Target size analysis by radiation inactivation is now a well-established method to study structure-function relationships in biologically active macromolecules without prior purification or even solubilization. Recently, it was reported that a relatively low-dose-rate but commonly available gamma source such as the Gammacell 220 (Atomic Energy of Canada, Ltd.) can be used to carry out radiation inactivation experiments providing it is appropriately calibrated with enzymes of known radiation sensitivities (G. Beauregard and M. Potier (1982) Anal. Biochem. 122, 379-384). In this report, a tube rack designed to fit into the irradiation chamber of the Gammacell 220 which allows five experiments (at 30 tubes per experiment) to be carried out simultaneously with both standard and unknown samples is described. The dose rates delivered at different positions in the rack were determined by irradiating rat liver cytosolic neuraminidase, an enzyme of known radiation sensitivity. A better than 2.7% agreement was obtained between experimental dose rate and computed values from isodose curves previously published by other authors (O. A. Curzio and H. O. Quaranta (1982) Int. J. Appl. Radiat. Isot. 33, 1-3).

Animals↗
PubMed1983

The target sizes of the in situ and solubilized forms of human placental steroid sulfatase as measured by radiation inactivation.

Whole microsomal membrane preparations and Triton X-100-solubilized human placental steroid sulfatase were subjected to radiation inactivation analysis using gamma rays from a 60Co irradiator in order to assess the size of the physiological form of the enzyme. The data indicate that the enzyme exists as a monomer of molecular weight 78600 in Triton-containing buffers and as a polymer of molecular weight 533000 within the microsomal membrane.

Female↗