Biomedical subjects

S Cortassa

Publications and source records attributed to S Cortassa.

28 records · Page 2Linked to original sources

PubMed1994

Spatio-temporal regulation of glycolysis and oxidative phosphorylation in vivo in tumor and yeast cells.

Recent advances in the in vivo control and regulation of glycolysis and oxidative phosphorylation in yeast and tumor cells is revised. New insights are presented from old and new experimental data interpreted in the light of powerful new technologies (e.g. NMR, confocal microscopy) and quantitative techniques combined with mathematical modeling. Those new aspects are mainly concerned with the dynamical organization of glycolysis and oxidative phosphorylation which emerges from the multiple interactions between compartments and processes inside the cells. Those compartments may be of structural origin, e.g. plasma membrane defining the cell boundary, mitochondrial-cytoplasmic, or functional ones such as the alternative association-dissociation of enzymes to subcellular structures (e.g. mitochondria, cytoskeleton) with different kinetic properties in each state. A novel regulatory mechanism concerning polymerization-depolymerization of microtubular protein may add a new dimension to the in vivo physiological properties of cells. One main suggestion coming from the modulatory power of the polymeric status and concentration of cytoskeleton components is that it could function as an intracellular mechanism of synchronization between microscopic (local) to macroscopic (global) processes. How the cell "mixes" or switches on or off those regulatory steps or effectors under different physiological and environmental conditions and for different genetic backgrounds, is a main avenue of systematic research for the future.

Animals↗

PubMed1993

Altered topoisomerase activities may be involved in the regulation of DNA supercoiling in aerobic-anaerobic transitions in Escherichia coli.

This study uncovers a new mechanism of regulation of DNA supercoiling operative in vivo upon an aerobic-anaerobic transition in Escherichia coli. Exponentially growing aerobic batch cultures were subjected to a shift to anaerobic conditions. The ratio [ATP]/[ADP] remained essentially constant at 8.5 in the aerobic culture and after a transition to anaerobiosis while DNA supercoiling increased noticeably upon anaerobiosis. This result indicated that the mechanism of regulation of DNA supercoiling by the [ATP]/[ADP] ratio was not operative. The increase in DNA supercoiling was followed by a large decrease in the DNA-relaxing activity of topoisomerase I while gyrase activity remained relatively constant. This decrease in the activity of topoisomerase I is likely to be responsible for the increase in DNA supercoiling.

Adenosine Diphosphate↗

PubMed1993

An allometric interpretation of the spatio-temporal organization of molecular and cellular processes.

Different levels of organization distinguished by characteristics spatial dimensions, Ec, and relaxation times, Tr, of biological processes ranging from electron transport in energy transduction to growth of microbial and plant cells, are shown to be related through a relation that may be interpreted as allometric and characterized by two different slopes. Processes, at levels of organization occurring in spatial dimensions of micrometers and relaxing in the order of minutes, delimit a 'transition point' between the two curves, that we interpret as a limit for the emergence of macroscopic coherence. The characteristic spatial dimension, Ec, and the relaxation time, Tr, contain dynamical information about the processes occurring at a given level of organization. When a steady state of a biological process at a certain level of organization becomes unstable, the system undergoes a transition to another level of organization. To exemplify the appearance of macroscopic order at levels of organization further from the 'transition point' we present in this report various experimental systems involving many levels of organization allometrically related that exhibit different kinds of self-organized behavior, i.e. bi-stability, oscillations, changes in (a)symmetry.

Cell Division↗

PubMed1992

Fractal organisation in biological macromolecular lattices.

Macromolecular solutions of proteic, glycoproteic or polysaccharidic nature in the presence of salt gave rise (when slowly dried) to dendritic-like fractal patterns. A fractal dimension D = 1.79 +/- 0.018 was obtained for dendritic patterns of a sonicated ovomucin gel (40% ovomucin-60% ovalbumin) in the presence of 0.1 M NaCl. The calculated D is similar to that described in percolation clusters. The appearance of fractal patterns was dependent upon the protein:salt ratio with an optimum in the range 0.75-1.25. Patterns disappeared at either lower or higher ratios. We conclude that the salt percolates through the macromolecular lattice and precipitates in fractal clusters during the drying process. Dendritic-like fractal patterns with similar D and morphologies were obtained with solutions of fetuin, ovalbumin, albumin or starch suggesting that fractal patterning is a general property of biological polymers. That cellular polymers would also aggregate in a fractal way was implied from the analysis of cellular cytoskeleton and microtrabecular lattices.

Glycoproteins↗

PubMed1991

Linear nonequilibrium thermodynamics describes the dynamics of an autocatalytic system.

A model simulating oscillations in glycolysis was formulated in terms of nonequilibrium thermodynamics. In the kinetic rate equations every metabolite concentration was replaced with an exponential function of its chemical potential. This led to nonlinear relations between rates and chemical potentials. Each chemical potential was then expanded around its steady-state value as a Taylor series. The linear (first order) term of the Taylor series sufficed to simulate the dynamic behavior of the system, including the damped and even sustained oscillations at low substrate input or high free-energy load. The glycolytic system is autocatalytic in the first half. Because oscillations were obtained only in the presence of that autocatalytic feed-back loop we conclude that this type of kinetic nonlinearity was sufficient to account for the oscillatory behavior. The matrix of phenomenological coefficients of the system is nonsymmetric. Our results indicate that this is the symmetry property and not the linearity of the flow-force relations in the near equilibrium domain that precludes oscillations. Given autocatalytic properties, a system exhibiting liner flow-force relations and being outside the near equilibrium domain may show bifurcations, leading to self-organized behavior.

Catalysis↗

PubMed1991

Dynamic regulation of yeast glycolytic oscillations by mitochondrial functions.

The control exerted in vivo by mitochondrial functions on the dynamics of glycolysis was investigated in starved yeast cells that were metabolizing glucose semianaerobically. Glycolytic oscillations were triggered after a pulse of glucose by inhibition of mitochondrial respiration with KCN, myxothiazol and antimycin A or in mutants in the bc1 complex (ubiquinol:cytochrome c reductase) that were largely deficient in respiratory capacity. Inhibition of the adenine nucleotide translocator by preincubation with bongkrekic acid also triggered a train of damped sinusoidal oscillations after glucose addition. The oscillations consisted of cycles of reduction and oxidation of the intracellular pool of nicotinamide nucleotides with periods of 45 s to 1 min and amplitudes of 0.8 mM or lower. Preincubation with the uncoupler carbonyl cyamide p-(trifluoromethoxy)phenylhydrazone (FCCP) annihilated cyanide-induced oscillations of NAD(P)H. Evidence for de-energization of mitochondrial membranes in vivo was obtained by mitochondrial staining with dimethylaminostyryl-methyl-pyridiniumiodine (DASPMI) of starved cells. The low rates of NADH reoxidation shown by respiratory mutants and the FCCP-treated X2180 strain open up the possibility that mitochondrial dehydrogenases also control glycolytic oscillations. Low rates of cytosolic NADH reoxidation induced by pyrazole, an inhibitor of alcohol dehydrogenase, were also associated with the disappearance of glycolytic oscillations. From experimental evidence and model calculations we conclude that the modulation of the levels of cytosolic ATP by mitochondrial functions in turn modulates the approach of the dynamic behavior of glycolysis to an oscillatory domain. The mitochondrial NADH dehydrogenase and the glycolytic steps associated with NADH reoxidation downstream from pyruvate appear to provide another control level of glycolysis dynamics in vivo.

Adenosine Triphosphate↗

PubMed1990

Pattern formation in an immobilized bienzyme system. A morphogenetic model.

Experimental and theoretical studies of a reaction-diffusion model of two immobilized enzymes participating in the cellular acid-base metabolism, namely glutaminase and urease, are presented. The system shows an unstable steady state at pH 6.0, where any perturbation will drive the system towards a more alkaline or more acidic pH, owing to the autocatalytic behaviour with respect to pH exhibited by both enzymes. When diffusion is coupled to reaction by means of immobilization, different patterns of the internal pH profile appear across the membrane. If the bienzymic membrane is subjected to a perturbation at its boundaries, of the same amplitude but in opposite directions, the internal pH evolves through an asymmetric pattern to attain a nearly symmetric distribution of pH. The pH value at the final steady state is more acidic or more alkaline than the initial state according to the initial and boundary conditions. The final nearly symmetric state is attained more rapidly when less enzyme is immobilized (1.8 x 10(-4) M.s-1 as against 3.3 x 10(-4) M.s-1 of total enzyme activity in the membrane volume). The experimental results agree rather well qualitatively with numerical predictions of the model equations.

Diffusion↗

PubMed1989

pH-induced bistable dynamic behaviour in the reaction catalysed by glucose-6-phosphate dehydrogenase and conformational hysteresis of the enzyme.

1. Bistable (multiple stationary states) dynamic behaviour in the activity of glucose-6-phosphate dehydrogenase that was subjected to successive pH change was demonstrated in an open continuously stirred tank reactor. Although the enzyme under study did not exhibit an autocatalytic effect and was homogeneously distributed, bistability was shown to occur. 2. The successive pH changes of the enzyme solution corresponded to a pH transition (8.3 in equilibrium 2), i.e. an acidification (forward direction) and an alkalinization (reverse direction). By use of intrinsic protein fluorescence methods, a glucose-6-phosphate dehydrogenase conformational hysteresis was shown to exist concomitant with the pH transition before and after enzyme injection into the reactor. 3. The results obtained suggest that the enzyme behaves, conformationally, as a memory device that stores information about its pH history (i.e. the enzyme records information in its structure about the environment to which it was previously exposed) and transduces it in a non-linear dynamic fashion, producing the bistable behaviour observed in the open reactor.

Catalysis↗

PubMed1989

Effect of phospholipids on the activity of sialosyl lactosylceramide (GM3): N-acetylgalactosaminyl transferase from chick embryo brain.

A preparation of UDP-GalNAc:sialosyl lactosylceramide N-acetylgalactosaminyl transferase (E.C. 2.4.1.92) obtained from 14-day chick embryo brain was delipidated partially by treatment with cold acetone in the presence of varying amounts of sodium dodecyl sulphate (SDS). The lipid content and the enzyme activity of the preparation decreased as the concentration of SDS increased. At 0.3% SDS the lipid content was about 30% and the enzyme activity about 15% of the original. The activity could be restored up to 60% of the original by added phospholipids, provided the removal of endogenous lipids did not exceed 70%. Phospholipids with different composition showed different abilities to restore the enzyme activity. Among phosphatidylcholines the decreasing order of effectivity was dilauroyl----dimiristoyl----dipalmitoyl----distearoyl-choline. Dimiristoyl phosphatidylcholine, dimiristoyl phosphatidylglycerol and dipalmitoyl phosphatidylglycerol activated the enzyme more effectively than dimiristoyl phosphatidyl ethanolamine, dimiristoyl phosphatidic acid or brain phosphatidylserine. No correlation was found between the activating ability and the charge in the polar head group of the lipid added. Addition of dilauroyl phosphatidylcholine to the delipidated preparation increased about 5-fold the Vmax without affecting the apparent Km for both the donor nucleotide and acceptor glycolipid. The data suggest that the lipid composition of the enzyme environment constitutes a potential level of regulation of the activity of this key enzyme of ganglioside biosynthesis.

Animals↗

PubMed1984

Biosynthesis of gangliosides in the developing chick embryo retina.

Chick embryo retinas were cultured with [3H]glucosamine on each day between days 6 and 12 of development. The total labeling of gangliosides decreased from day 6 to day 12. The decrease was mostly due to the decrease in the labeling of disialosyllactosylceramide (GD3), which diminished to less than 2%, while the labeling of the disialoganglioside GD1a decreased to about 30%. Labeling of endogenous gangliosides by incubating retina homogenates with CMP-[3H]N-acetylneuraminic acid, in addition to showing a decrease in the labeling of GD3, showed a twofold increase in the labeling of GD1a. The changes in the pattern of labeling correlated with the decrease of both the incorporation of [3H]thymidine into DNA and the activity of CMP-NeuAc:GM3 sialosyltransferase and with the increase of both the activities of UDP-Ga1NAc:GM3 N-acetylgalactosaminyltransferase and choline acetyltransferase. The results suggest that the shift in the pattern of labeling of gangliosides occurs in association with the transition from the proliferative to the nonproliferative state of neuronal cells. Retinas at each age cultured with [3H]glucosamine for 22 hr maintain the radioactivity in gangliosides for at least 4 additional days in culture. This indicates that within this developmental interval there is not a period characterized by having a high turnover rate of gangliosides.

Age Factors↗