Search PubMed⌕ Search

Biomedical subjects

M T Debanne

Publications and source records attributed to M T Debanne.

34 records · Page 2Linked to original sources
PubMed1980

Multivalent interaction between asialofetuin and plasma membrane preparations from the rat liver.

Binding of bovine asialofetuin by rat liver plasma membranes was studied using different techniques for the separation of the free and bound forms of the glycoprotein and also different approaches to measure nonspecific binding. The membrane preparations had the electron microscopic appearance of a mixture of lamellae and vesicles and their lipid:protein ratios and marker enzyme profiles fell within the range of values available from the literature. The binding capacity was approximately 15 pmol of asialofetuin per milligram of membrane protein. Scatchard plots of the values obtained over a wide range of concentrations (4.8--12.6 micrograms asialofetuin per 30 micrograms membrane protein) after incubation at 22 degrees C showed pronounced nonlinearity which, in combination with evaluations according to other theoretical models, was referable to heterogeneity of binding. In sharp contrast, after incubation at 4 degrees C the Scatchard plot was linear. This difference is interpreted as the expression of a functional, rather than a chemical, heterogeneity in asialofetuin binding. The underlying mechanism is thought to be competition of galactose groups for binding sites with the result that the number of bonds varies between the galactose groups of a bound asialofetuin molecular and the hepatic lectin, depending on the concentration of the glycoprotein in the incubation mixture.

Animals↗
PubMed1979

Three types of human asialo-transferrin and their interactions with the rat liver.

Three types of asialo-transferrin were obtained from immunologically pure human transferrin by chromatography on DEAE-cellulose, followed by desialylation and affinity chromatography on a column of the immobilized asialo-glycoprotein-binding hepatic lectin from rabbit liver. Of the asialo-transferrins, type 1 was derived from the principal DEAE-cellulose chromatographic component of transferrin, i.e. the one that contains two biantennary glycans. The two other asialo-transferrins (types 2 and 3) were derived from a minor DEAE-chromatographic transferrin component, which is assumed to possess one biantennary and one triantennary glycan. The three asialo-transferrin types were indistinguishable by electrophoretic mobility, but they were readily distinguished on the basis of their binding strengths to the hepatic lectin in intact rats. Glycan structures responsible for the difference in binding strengths between asialo-transferrin types 2 and 3 are not known. Metabolic studies in rats showed that none of the individual asialo-transferrin types was capable of generating a signal for endocytosis at low doses (<1mug/100g body wt.) and, consequently, most of the injected protein was recoverable with the plasma and the liver 35min after injection. However, endocytosis and catabolism of each asialo-transferrin type was readily induced by injecting a larger dose (50-250mug/100g body wt.) of unlabelled asialo-transferrin of the same type or of a different type a short interval after the labelled dose. These findings support the view that the dose-dependent uptake of human asialo-transferrin by the hepatocyte, as established in an earlier study with asialo-transferrin made from whole transferrin [Regoeczi, Taylor, Hatton, Wong & Koj (1978) Biochem. J.174, 171-178], also holds for these asialo-transferrin subfractions. Furthermore, the present studies indicate that asialo-transferrins of different carbohydrate compositions are capable of synergistically promoting endocytosis of each other.

Animals↗
PubMed1979

Bi-and tri-antennary human transferrin glycopeptides and their affinities for the hepatic lectin specific for asialo-glycoproteins.

Glycopeptides were isolated from a proteolytic digest of human transferrin. After mild acid hydrolysis the desialylated glycopeptides were labelled by the galactose oxidase/NaB(3)H(4) procedure and then fractionated by Sephadex-gel filtration or by anion-exchange chromatography. Either technique allowed separation of the two heterosaccharide chains (designated glycan I and glycan II) previously described for this protein by Spik, Vandersyppe, Fournet, Bayard, Charet, Bouquelet, Strecker & Montreuil (1974) (in Actes du Colloque Internationale No. 221 vol. 1, pp. 483-499). Subsequent chromatography on Sepharose-concanavalin A separated fractions containing different quantities of carbohydrates for each glycan, as indicated by analyses. The isolated glycan fractions were then tested for their abilities to bind to the immobilized rabbit hepatic lectin. Our studies suggest that either glycan can have a bi- or tri-antennary structure. Desialylated biantennary glycans I and II did not bind to the hepatic lectin. Desialylated triantennary glycan I was slightly retarded by the hepatic lectin, whereas the triantennary glycan II consisted of equal quantities of a retarded and a bound type. Desialylated triantennary glycan II was totally displaced from the hepatic lectin by using a buffer containing 0.05m-EDTA. The results suggest that greater structural heterogeneity exists in the carbohydrate moiety of human transferrin than was previously envisaged. Such heterogeneity could be reflected in several molecular forms of human transferrin, which, after desialylation, differ significantly in their affinities for the hepatic lectin.

Amino Acids↗
PubMed1978

Elimination of asialofetuin and asialoorosomucoid by the intact rat. Quantitative aspects of the hepatic clearance mechanism.

The capacity of the liver to eliminate asialofetuin and asialoorosomucoid was investigated in intact rats. From plasma radioactivity curve measurements and assays on tissue homogenates the liver is shown to be able to dispose of an average of 19.8 microgram of asialofetuin/min per 100 g body weight. No other major route is identified for the disappearance of asialofetuin from the plasma, although trace amounts of the protein were detectable in the urine. From analyses of the plasma radioactivity curves the elimination process for asialoorosomucoid appears to be comparatively complex because of the existence of extrahepatic disposal routes. Quantification of labelled asialoorosomucoid in liver homogenates indicates, however, that the hepatic clearance rate for asialoorosomucoid is similar to that for asialofetuin. Urinary excretion significantly contributes to the disappearance of asialoorosomucoid from the plasma but the hepatic and renal routes do not account for all the protein lost from this compartment. At plasma concentrations above the maximal eliminative capacity of the liver, the hepatic clearance of asialofetuin obeys zero-order kinetics and is remarkably constant. Elimination of a quantity of asialoglycoprotein which exceeds the calculated total number of binding sites in the liver does not reduce the efficiency of the pathway, and studies of [3H]leucine incorporation indicate that the lectin, unlike the bound asialoglycoprotein, is not destroyed in the elimination process. Cytochalasin B (80 microgram/100 g body wt.) had no measureable effect on the hepatic clearance of asialofetuin. Administration of colchicine (10 mg/100 g body wt.) resulted in transitory accumulations of asialoorosomucoid in the liver, presumably due to interference with the intracellular transport of the endocytised protein.

Animals↗
PubMed1976

Characteristics of the macrophage uptake of proteinase-alpha-macroglobulin complexes.

Complexes formed between labelled proteolytic enzymes (trypsin, subtilopeptidase A) and the alpha-macroglobulins of plasma are rapidly and selectively taken up by rabbit alveolar macrophages. The uptake occurs over a narrow zone of pH. Kinetics of the uptake is affected by temperature; in particular, incubation of macrophages at 37 degrees C before the addition of the labelled complex reduces the capacity to take up complexes. EDTA prevents the association of labelled complexes with macrophages, and can dissociate previously bound label. The effect of EDTA is reversed by the addition of calcium or magnesium or both. Iodoacetamide does not prevent the uptake of complexes but causes them to remain available for dissociation from the cells by EDTA. Incubation of complexes with macrophages at 37 degrees C with no iodoacetamide results in the appearance of trichloroacetic acid soluble products of the enzyme in the supernatant fluid. These observations indicate that the selective uptake of proteinase-alpha-macroglubin complexes by rabbit alveolar macrophages can be resolved into three phases: (1) membrane binding which depends upon divalent cations and is pH sensitive, (2) endocytosis inhibitable by iodoacetamide and (3) temperature-dependent hydrolysis of the contained labelled enzyme.

Animals↗
PubMed1975

Uptake of proteinase-alpha-macroglobulin complexes by macrophages.

Complexes of labelled proteinases (subtilopeptidase A, trypsin) with serum alpha 1-macroglobulin or alpha 2-macroglobulin are rapidly taken up in vitro by rabbit alveolar macrophages and peritoneal macrophages but not by mixed rabbit peripheral blood leukocytes. Enzyme, not bound to alpha 1- or alpha 2-macroglobulin, does not become associated with alveolar macrophages. Chemically inactivated subtilopeptidase A does not bind to alpha 1- or alpha 2-macroglobulin; chemically inactivated subtilopeptidase A in mixtures with alpha 1 - or alpha 2-microglobulin, does not interact with alveolar macrophages. Blocking experiments confirmed that the interaction of proteinase with alveolar macrophages is complex specific; uptake of labelled complex was prevented by the simultaneous addition of macroglobulin complexes formed with non-labelled subtilopeptidase A, subtilopeptidase B, trypsin or chymotrypsin but not by macroglobulin alone. The findings demonstrate a complex-specific interaction between proteinase-alpha-macroglobulin complexes and macrophages.

Animals↗
PubMed1975

The role of alpha1-antitrypsin and alpha-macroglobulins in the uptake of proteinase by rabbit alveolar macrophages.

Proteinases in plasma bind largely to alpha1-antitrypsin and alpha-macroglobin. The latter is represented in humans by alpha2-macroglobulin and in the rabbit by alpha2-macroglobulin and alpha1-macroglobulin. There is a selective and rapid uptake of proteinase-macroglobulin complexes by rabbit alveolar macrophages. It was found that complexes of labelled trypsin or subtilopeptidase A with rabbit alpha1-antitrypsin do not become similarly associated with rabbit alveolar macrophages. Moreover, proteinase-alpha1-antitrypsin complexes failed to inhibit the uptake of labelled proteinase-macroglobulin complexes. Thus, the interaction with macrophages of proteinase-macroglobulin complexes represents a pathway of proteinase metabolism distinct from that involving proteinase bound to alpha1-antitrypsin.

Animals↗
PubMed1974

Effect of nitrate on the synthesis and decay of nitrate reductase of Neurospora.

1. A method was developed to examine the turnover of nitrate reductase by the use of tungstate. 2. Evidence is presented which suggests that the disappearance of nitrate reductase activity from Neurospora mycelia exposed to non-inducing conditions is due to the disappearance of the enzyme protein(s) from the mycelia, and not merely due to the disappearance of its (their) catalytic power. 3. The presence of NO(3) (-) in the culture medium slows down the rate of degradation of nitrate reductase in Neurospora in vivo.

Animals↗
PubMedDate not supplied

Transferrin glycans: a possible link between alcoholism and hepatic siderosis.

The hepatic uptake of 59Fe from diferric rat and rabbit asialotransferrins and from human transferrin lacking two sialyl residues was investigated in rats in experiments lasting for 1 hr. The 59Fe attached to either of these preparations disappeared from the plasma more rapidly than the 59Fe introduced with the unmodified respective parent proteins. Most of the 59Fe activity that had disappeared from the circulation could be recovered with the liver. Studies with double-labeled (125I, 59Fe) preparations showed that the enhanced 59Fe clearance was not associated with increased catabolism of the modified transferrins. Prolonged, heavy alcohol consumption, as shown by others, results in the appearance of sialic acid-deficient transferrin (two residues missing) in human serum. We suggest that the increased capacity of transferrin deficient in sialic acid to selectively deposit iron in the hepatocyte may be of significance for the development of the hepatic siderosis observed in alcoholism.

Alcoholism↗