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R Colonna

Publications and source records attributed to R Colonna.

At least 55 records · Page 3Linked to original sources
PubMed1987

Lipid interaction of diphtheria toxin and mutants with altered fragment B. 1. Liposome aggregation and fusion.

The interaction of diphtheria toxin and its cross-reacting mutants crm 45,228 and 1001 with small unilamellar vesicles has been followed by a turbidity assay, electron microscopy, fluorescence energy transfer and membrane permeability. All toxins at pH lower than 6 induce the aggregation and fusion of liposomes containing negatively charged phospholipids; crm 45 and crm 1001 are less potent than diphtheria toxin. Isolated diphtheria toxin fragment B is very effective while isolated fragment A is ineffective. Liposome fusion induced by the toxins at low pH occurs without release of the internal content implying that fusion does not involve vesicle breakage and resealing. The pH dependence of the membrane interaction of diphtheria toxin monitored by turbidity is in close agreement with that monitored by fluorescence energy transfer. It shows that diphtheria toxin can alter the lipid bilayer structure in the pH interval 5-6. This pH range occurs in endosomes and suggests that histidyl and carboxyl residues are likely to be involved in the conformational change of diphtheria toxin triggered by acidic pH.

Diphtheria Toxin↗
PubMed1987

Diphtheria toxin and its mutant crm 197 differ in their interaction with lipids.

The interaction of diphtheria toxin and its enzymatically deficient mutants crm 176 and crm 197 with liposomes has been studied by turbidity measurement and hydrophobic photolabelling with photoactivatable phosphatidylcholines. Diphtheria toxin and crm 176 at neutral pH bind to the surface of lipid bilayers while crm 197 also appears to interact with the fatty acid chains of phospholipids. All proteins undergo a change in conformation over the same range of acidic pH and become able to insert in the lipid bilayer. The tighter lipid interaction of crm 197 may account for its higher cell association constant. The possibility is discussed that the binding of diphtheria toxin to cells is mediated by both a protein receptor and an interaction with the head group of phospholipids.

Diphtheria Toxin↗
PubMed1987

Follicle cell regulation of mammalian oocyte growth.

To investigate mechanisms of follicle cell control on mammalian oocyte growth, preantral mouse oocytes free from surrounding follicle cells were individually cocultured with monolayers of different somatic cells competent to form gap junctions, and the rate of in vitro oocyte growth was directly correlated with the level of metabolic coupling on the same cells. The results indicate that 1) at a similar extent of metabolic coupling, mouse oocytes grew on follicle cells but not on 3T3 and Sertoli cell monolayers, and 2) the growth rate of oocytes cultured on follicle cells was dependent on the extent of metabolic coupling. It was concluded that gap-junction-mediated nutrition of ovarian mouse oocytes exerted by somatic cells is necessary but not sufficient to maintain oocyte growth. A specific regulatory role of follicle cells on mammalian oocyte growth is proposed.

Animals↗
PubMed1986

Lipid miscibility and size increase of vesicles composed of two phosphatidylcholines.

The size increase of small unilamellar vesicles composed of binary mixtures either of saturated fatty acid phosphatidylcholines with different chain lengths or of saturated and unsaturated phosphatidylcholines was found to depend on the miscibility properties of the lipid components. No size increase was detected in vesicles formed by two miscible phosphatidylcholines. In vesicles composed of two lipids which are partially immiscible in the gel state, a size increase was observed at temperatures which mainly overlapped the range of temperatures of the lipid phase transition. The rate of size increase of vesicles composed of two lipids which are immiscible in the gel state was faster than that of vesicles composed of two partially immiscible phosphatidylcholines, and the process occurred not only at the temperature ranges of the lipid phase transition, but also when both lipids were in the gel state. The vesicle size increase process occurred without the mixing of the internal content of the vesicles. A model is proposed in which the presence of 'fractures' between membrane regions of different fluidity and/or lipid composition controls the rate of this process.

Chemical Phenomena↗
PubMed1986

Gramicidin induced aggregation and size increase of phosphatidylcholine vesicles.

To investigate the role of membrane proteins in the fusion process, linear hydrophobic polypeptide gramicidin was used as fusogenic agent in small unilamellar vesicles (SUV) constituted of saturated lecithins. It was found that gramicidin, externally added to a suspension of vesicles, induces a reversible vesicles aggregation. When incorporated into the bilayer, gramicidin induces increase in vesicle size. The vesicle size increase was monitored by column chromatography and transmission electron microscopy. The process of vesicle size increase occurs only when the lipid membrane is in the gel state. A maximum is observed in the kinetics at a temperature of approx. 25 degrees C lower than the phase transition temperature of lipids. Higher rates of vesicle size increase are obtained as the lipid chain length increases. The process is accompanied by a release of internal vesicle content and by membrane lipid mixing.

Brain↗
PubMed1984

Stage-dependent modifications of amino acid uptake by antral and metaphase II mouse oocytes.

Modifications of leucine transport system of mouse oocytes have been studied throughout Graafian follicle development and oocyte maturation. In contrast to sheep oocytes (Moor and Smith, 1979), in the mouse kinetic constants and efflux rate of leucine transport system did not vary in diestrus, proestrus, and metaphase II (met II) oocytes. However, kinetics of leucine equilibration in proestrus and met II oocytes was significantly slower than that found in diestrus cells, and this may reflect a decreased availability of internal amino acids for exchange.

Animals↗
PubMed1983

Voltage oscillations in mammalian metaphase II oocytes.

The membrane potential has been measured in ovulated mouse oocytes using conventional electrophysiological techniques. Temporal oscillations in membrane voltage have been observed in the oocytes, with periods of about 6 h. This oscillatory pattern, peculiar to oocytes in metaphase II, might explain the differences in membrane potential values reported in several studies on mammalian oocytes.

Animals↗
PubMed1983

Amino acid transport systems in growing mouse oocytes.

Amino acid transport systems have been studied in isolated mouse oocytes throughout oogenesis. While these cells were lacking the A-transport system of Ehrlich cell (Christensen, 1975), they were found to be provided with the L- and ASC-systems of Ehrlich cell (Christensen, 1975). Since, in contrast to the A-system, the L- and ASC-systems mainly work by exchanging internal and external amino acids, we propose that their main function is to actively maintain the appropriate internal balance between different amino acids within the growing oocyte.

Alanine↗
PubMed1983

Mechanisms of amino acid uptake in cumulus-enclosed mouse oocytes.

The nutritional role of mouse granulosa cells on antral dictyate mouse oocytes has been studied by measuring the transfer of different amino acids through gap junctional channels between somatic and germ cells. When present in the incubation medium at concentrations resembling in vivo conditions, glycine, alanine, proline, serine, tyrosine, glutamic acid and lysine entered cumulus-enclosed oocytes cooperatively, while valine, leucine and phenylalanine did not. However, cooperative uptake of leucine and phenylalanine was observed at higher external precursor concentrations. We conclude that in vivo antral mouse oocytes depend on surrounding granulosa cells for amino acid uptake, with the exception of amino acids carried by the leucine exchange transport system, and propose that amino acid transfer between granulosa cells and oocytes is dependent on precursor concentrations in the coupled cells.

Amino Acids↗
PubMed1982

Effect of lipid mixing on the permeability and fusion of saturated lecithin membranes.

The effect of phospholipid mixing on the permeability properties of multilamellar lipid vesicles (MLV) was studied. In the solid state, dimyristoylphosphatidylcholine/dipalmitoylphosphatidylcholine (DMPC/DPPC) vesicles exhibit ideal lipid miscibility; dimyristoylphosphatidylcholine/distearoylphosphatidylcholine (DMPC/DSPC) vesicles exhibit nonideal lipid miscibility at low DSPC molar fractions; dimyristoylphosphatidylcholine/dibehenoylphosphatidylcholine range of DBPC molar fractions. The rates of K+, ethylene glycol, and water diffusion from these vesicles in the solid state were measured by photometric and electrometric techniques. The following results were obtained: (1) The rate of solute diffusion, which is decreased monotonically, in DMPC/DPPC MLV, by increasing the molar fractions of DPPC, exhibits maxima at 0.2 molar fraction of DSPC in DMPC/DSPC MLV and at 0.4 molar fraction of DBPC in DMPC/DBPC MLV. (2) The activation energy of the solute diffusion process abruptly decreases in approximately the same range of lipid molar fractions where nonideal lipid miscibility is present. (3) The membrane pore radius is increased by increasing the lipid nonideal miscibility. The rate of vesicle size increase, measured by absorbance changes, is decreased monotonically in DMPC/DPPC monolamellar vesicles [small unilamellar lipid vesicles (SUV)] by increasing the molar fraction of DPPC. It, however, exhibits a maximum in DMPC/DSPC SUV at 0.15 molar fraction of DSPC. A model was suggested in which the solute diffusion and the membrane fusion processes are controlled by fractures. The average width of the fractures is increased by increasing the lipid immiscibility.

Diffusion↗
PubMed1980

Phospholipid release from sonicated vesicles induced by a "critical" fatty acid concentration.

Dipalmitoyl phosphatidylcholine vesicles incubated in the presence of increasing amounts of myristic acid showed a progressive translocation of phospholipid molecules across a dialysis membrane. The rate of phospholipid translocation increased abruptly at a 'critical' value of myristic acid concentrations. The translocation rate of mixed dipalmitoyl phosphatidylcholine/myristic acid vesicles obtained by cosonicating the two components was also dependent on a 'critical' fatty acid concentration. A marked release of K+ and different responses of fluorescent probes to the fatty acid addition were observed at this concentration.

Chromatography, Gel↗
PubMed1980

Phospholipid exchange and size enlargement in sonicated vesicles induced by a 'critical' fatty acid concentration.

Size enlargement of dipalmitoyl phosphatidylcholine vesicles was greatly accelerated in the range of the phase-transition temperatures, when fatty acid concentration was above a threshold level ('critical' concentration). This 'critical' concentration varied with the length of the fatty acid chain. The size enlargement process had second-order kinetics dependent on the vesicle concentration. Alkaline pH and low ionic strength inhibited the rate of size enlargement. Phospholipid exchange between dimyristoyl and dipalmitoyl phosphatidyl-choline vesicles increased abruptly above a 'critical' fatty acid concentration. The donor vesicles were those vesicles in which fatty acids reached the 'critical' concentration. The phospholipid exchange occurred both in fluid- and in solidstate vesicles. The 'critical' fatty acid concentration accelerating the phospholipid exchange process was lower than that accelerating the size enlargement process. The phospholipid exchange process explained in terms of a diminished hydrophobic attraction among the phospholipid molecules of the bilayer occurs via a free phospholipid molecule transfer through the aqueous phase. The size enlargement process is interpreted in terms of high fatty acid concentration in the membrane fluid domains. The membrane structure is locally perturbed inducing vesicle sticking after collision.

Fatty Acids↗
PubMed1978

Interaction of cytochrome c with cytochrome c oxidase. Photoaffinity labeling of beef heart cytochrome c oxidase with arylazido-cytochrome c.

Cytochrome c derivatives labeled with a 3-nitrophenylazido group at lysine 13, at lysine 22, or at both residues have been prepared. The interaction of the cytochrome c derivatives with beef heart cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) in the presence of ultrviolet light results in formation of a covalent complex between cytochrome c and the oxidase. Using the lysine 22 derivative, the polypeptide composition of the oxidase is not modified, nor is its catalytic activity, whereas with the lysine 13 derivative, the gel electrophoretic pattern is altered and the catalytic activity of the complex diminished. The data are consisten with a specfic covalent interaction of the lysine 13 derivative of cytochrome c with the polypeptide of molecular weight 23,700 (Subunit II) of cytochrome c oxidase.

Affinity Labels↗
PubMed1977

Cation transport in cytochrome oxidase reconstituted vesicles.

Cation translocation across the membrane of cytochrome oxidase reconstituted vesicles may be followed with a simple spectrophotometric method. Cytochrome oxidase reconstituted vesicles, supplemented with ascorbate and cytochrome c. induce large spectral changes of the positive dye safranine, reversed by uncouplers and inhibitors of respiration. The dye is probably accumulated in the inner space of the vesicles, where it reaches high concentrations and aggregates. The spectral shifts and the absorbance changes, due to aggregation, are proportional to the amount of the dye taken up and depend on the respiratory control. In the presence of potassium, valinomycin causes an inhibition, whereas nigericin stimulates the dye uptake. The data are discussed in terms of electrical potential dependent fluxes.

Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone↗