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M Markus

Publications and source records attributed to M Markus.

At least 37 records · Page 2Linked to original sources
PubMed1995

The organelles containing porcine 17 beta-estradiol dehydrogenase are peroxisomes.

Porcine 17 beta-estradiol dehydrogenase was recently purified and cloned. It catalyzes the NAD(+)-dependent oxidation of estradiol to estrone 360-fold more efficiently than the reverse reaction with NADPH. Immunogold electron microscopy localizes 17 beta-estradiol dehydrogenase in organelles of 120 to 500 nm with moderate electron-dense matrices bounded by single membranes. Antibodies against the peroxisomal markers catalase and acyl-CoA oxidase recognize the same organelles in double-labeling studies. This is the first report on the participation of peroxisomes in the metabolism of estradiol.

Acyl-CoA Oxidase↗
PubMed1994

Bumetanide and furosemide inhibited vascular endothelial cell proliferation.

In this study, we examined the role of the bumetanide-sensitive Na+/K+/Cl-cotransport in the mitogenic signal of vascular endothelial cell proliferation. The activity of the Na+/K+/Cl- cotransport is dramatically decreased in quiescent subconfluent cells, as compared to subconfluent cells growing in the presence of FGF. The Na+/K+/Cl- cotransport activity of quiescent subconfluent cultures deprived of FGF decreased to 6%, whereas that of quiescent cells grown to confluency was reduced to only 33% of the activity of subconfluent cells growing in the presence of FGF. The basal low activity of Na+/K+/Cl- cotransport in the quiescent subconfluent vascular endothelial cells was dramatically stimulated by FGF. In order to explore the role of the Na+/K+/Cl- cotransport in the mitogenic signal of the endothelial cells, the effect of two specific inhibitors of the cotransport -furosemide and -bumetanide was tested on cell proliferation induced by FGF. Bumetanide and furosemide inhibited synchronized cell proliferation measured by direct counting of cells and by DNA synthesis. Inhibition by furosemide and bumetanide was reversible; removal of these compounds completely released the cells to proliferate. These results indicate that the effect of these drugs is specific and is not due to an indirect toxic effect. This study clearly demonstrates that the FGF-induced activation of the Na+/K+/Cl- cotransport plays a role in the mitogenic signal pathway of vascular endothelial cells.

Animals↗
PubMed1994

[Risk factors in hyperlipidemic patients with coronary heart disease one year after PTCA].

The reduction of modifiable risk factors in hyperlipidemic patients with coronary heart disease (CHD) receiving standard medical care in Germany has not been evaluated before. We identified all patients < 65 yrs of age with marked hyperlipidemia (Cholesterol (Chol) > 250 mg/dl, HDL < 20% Chol) among all patients who underwent PTCA during defined periods 1991/92 in the Cardiology department of the Heinrich-Heine-University. The study patients (n = 93, age 54 +/- 8 yrs, 75 men, 18 women) were evaluated 13 +/- 2 months after PTCA for modification of their risk factors, treatments, knowledge, medications and dietary habits with a structured questionnaire and a 5-day dietary protocol. At follow-up Chol and LDL were significantly reduced from 299 +/- 47 to 253 +/- 43, and from 228 +/- 47 to 189 +/- 42 mg/dl, respectively (p < 0.001). 10% of patients had an LDL < 135 mg/dl. In contrast, HDL (43 +/- 9 mg/dl) and triglycerides (242 +/- 138 mg/dl), the percentage of smokers (40%), of overweight patients (38%) and of patients with elevated blood pressure (43%) remained unchanged. During the observation period the number of contacts with physicians was high (12 (1-40)). 32% of patients participated in dietary counselling by a dietician and 42% in an in-patient rehabilitation programme. In 2/3 of patients the knowledge related to hyperlipidemia and a lipid-lowering diet was good. However, according to the dietary protocols the fat intake was high (37 +/- 7% of total calories). Lipid lowering drugs had been prescribed in 68% of patients, predominantly as monotherapy and in low dosage.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Kinetics of inhibition of purified and mitochondrial cytochrome c oxidase by psychosine (beta-galactosylsphingosine).

1. Psychosine (beta-galactosylsphingosine) is the toxic agent in Krabbe's disease (globoid cells leukodystrophy). It inhibits purified bovine heart mitochondrial cytochrome c oxidase; there is a rapid phase of inhibition (complete within 10-15 s) and a slower phase (complete within 10-15 min). Both phases are also seen in rat liver mitochondria. IC50 is about 200 microM psychosine in the purified enzyme and less than 20 microM in mitochondria. Psychosine inhibition is due to binding to cytochrome oxidase, not cytochrome c. 2. Bovine heart submitochondrial particles show inhibition similar to rat liver mitochondria. However, although proteoliposomes containing bovine heart cytochrome oxidase show an identical fast phase, they have no noticeable slow phase of inhibition. Addition of phospholipid liposomes to submitochondrial particles relieved the majority of psychosine inhibition, consistent with the removal of those molecules binding in the slow phase. Psychosine can inhibit cytochrome oxidase molecules facing in either direction in proteoliposomes and submitochondrial particles, suggesting that it can rapidly interact with both sides of a membrane when added externally. 3. At high ionic strength, the presence of psychosine decreases the Vmax. of cytochrome oxidase with little effect on the Km for cytochrome c. This non-competitive inhibition suggests that the psychosine-enzyme complex is kinetically inactive and not labile over the time course of the assay. Psychosine does not inhibit the reduction of haem a or haem a3 by artificial electron donors, but does inhibit the reduction of haem a by cytochrome c.

Animals↗
PubMed1991

Phorbol ester TPA inhibits the stimulation of bumetanide-sensitive Na+/K+/Cl- transporter by different mitogens in quiescent BALB/c 3T3 mouse fibroblasts.

In this study we examined the effect of the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on the bumetanide-sensitive Na+/K+/Cl- transporter in quiescent BALB/c 3T3 cells. We have shown that exposure of quiescent BALB/c 3T3 cultures to phorbol ester did not inhibit the basal bumetanide-sensitive Rb+ influx or efflux. In fact, at high concentration (100 ng/ml), TPA slightly stimulated the bumetanide-sensitive Rb+ influx and efflux. However, when the quiescent cultures were stimulated by serum or by defined growth factors, the stimulated fraction of the bumetanide-sensitive Rb+ influx was drastically inhibited by exposure of the cells to the phorbol ester TPA. Based on the above findings, we propose that activation of protein kinase C by the phorbol ester TPA does not inhibit the Na+/K+/Cl- cotransport activity; however it does suppress only the growth-factors-stimulated fraction of the cotransport in quiescent BALB/c 3T3 cells. These data propose that activation of kinase C has a regulatory feedback effect on the stimulation of the Na+/K+/Cl- cotransport activity by growth factors.

Animals↗
PubMed1991

Na+/K+/Cl- cotransport is stimulated by a Ca(++)-calmodulin-mediated pathway in BALB/c 3T3 fibroblasts.

In the present study, we investigated the role of intracellular Ca++ in the stimulation of the Na+/K+/Cl- cotransport in synchronized BALB/c 3T3 cells. The Na+/K+/Cl- cotransport was stimulated by the growth factors EGF, TGF-alpha, IGF-1, and IGF-2, which do not activate protein kinase C, but do induce a transient increase in free cytoplasmic Ca++. In addition, direct activation of protein kinase C by the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) did not affect the Na+/K+/Cl- cotransport activity of quiescent cells. The Na+/K+/Cl- cotransport was also stimulated by the above mitogens in cells pretreated with the phorbol ester TPA. This treatment led to a progressive decline in the activity of cellular protein kinase C. This result implies that cells deficient in protein kinase C may still support stimulation of the Na+/K+/Cl- cotransport. Taken as a whole, these findings suggest that the Na+/K+/Cl- cotransport is stimulated predominantly by a protein kinase C-independent mechanism in BALB/c 3T3 fibroblasts. Both the intracellular Ca++ antagonist 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) and two potent calmodulin antagonists, trifluoperazine (TFP) and chloropromazine (CP), blocked serum- and mitogen-stimulated Na+/K+/Cl- cotransport. These results suggest that the Na+/K+/Cl- cotransport is stimulated by an increase of intracellular Ca++ and subsequently by a Ca(++)-calmodulin-mediated pathway in the synchronized BALB/c 3T3 fibroblasts.

3T3 Cells↗
PubMed1990

Modelling morphogenetic processes in excitable media using novel cellular automata.

As an efficient alternative to partial differential equations, cellular automata have been proposed for the simulation of excitable media. However, the automata proposed so far are anisotropic since the shapes of the propagating waves are related to the shapes of the cells (e.g. squares or hexagons). This problem is solved in the present work by automata based on a random distribution of excitable elements. The following results were obtained using a (minimal) model with only three parameters: a) Target patterns (concentric circular waves) and spirals in two dimensions in agreement with measurements in the Belousov-Zhabotinskii (BZ) reagent and in heart muscle during ventricular fibrillation. b) Waves rotating around a hole in agreement with observations on the cerebral cortex. c) A linear eikonal equation relating the normal velocity to the curvature of the wavefront in agreement with measurements in the BZ reagent and in the aggregating slime mold D. discoideum. d) Turbulences similar to those observed in experiments. e) Three-dimensional waves (scroll waves), some of which have been observed in thick heart muscle and in the BZ reagent. An extension of the model including two more parameters yields a dispersion relation comparable to that obtained from measurements and from an analysis of the relevant partial differential equations.

Animals↗
PubMed1987

On the recognition of order and disorder.

We compare several algorithms for the recognition of ordered and disordered images. As image sources we use waves of the Belousov-Zhabotinskii reaction coupled to convective motion in a petri dish. This device allows reversibly the generation of periodic (ordered) and aperiodic (disordered) patterns. The best match between the parametric description and the observations is obtained by an "autodifference function". This function is computed by summing up intensity differences over all pairs of picture elements having a given distance on the picture plane. Then, the minimum of this function is determined upon variation of the distance. This algorithm is not only efficient for the recognition of order and disorder in "machine vision", but also plausible in biological visual perception.

Form Perception↗
PubMed1985

Properties of strange attractors in yeast glycolysis.

The properties of periodic and aperiodic glycolytic oscillations observed in yeast extracts under sinusoidal glucose input were analyzed by the following methods. (1) Spectral analysis, rendering sharp peaks for periodic responses and enhanced broad-band noise for aperiodic oscillations. (2) Phase plane analysis, leading to closed and to open trajectories for periodic and aperiodic oscillations, respectively. (3) Rotation of a phase plane proportionally to time, revealing strange attractors associated with the aperiodic oscillations. (4) Stroboscopic plot on the phase plane, showing that the strange attractors follow a stretch-fold-press process, if the stroboscoping phase is varied. (5) Stroboscopic transfer plot, admitting a period of three transfer processes and thus implying chaos according to the Li-Yorke theorem. (6) Determination of the rate of information production by differentiation of the transfer plot, yielding approx. 0.21 bits per min for the chaotically glycolyzing yeast extract.

Journal Article↗
PubMed1985

Input-response relationships in the dynamics of glycolysis.

The dynamic response of glycolysis is investigated using a two-enzyme model with three different type of input flux time dependences: constant, sinusoidal, and amplitude or frequency modulated (AM or FM) input flux. The analysis reveals that the system acts as a device which transduces amplitudes and frequencies of the input function into complex and remarkably diverse response patterns. This device may have more than one response possibilities for a single input function, owing to the coexistence of attractors in phase space. In such a case one response function can be switched over into the other by short substrate pulses. Stroboscopic plots reveal that chaotic regimes obtained under sinusoidal input flux obey a stretch-fold-press process, similar to the baker's transformation, upon a change of the stroboscoping phase. Chaotic oscillations obtained under AM or FM conditions often show a much higher degree of randomness than those obtained under sinusoidal input. This is expressed quantitatively by an enhanced Liapunov dimension of the attractors.

Glycolysis↗
PubMed1984

Chaotic dynamics in yeast glycolysis under periodic substrate input flux.

The numerical analysis for a glycolytic model containing the enzymes phosphofructokinase and pyruvate kinase reveals different types of entrainment, as well as chaotic response under sinusoidal substrate input. Entrainment with response periods 1, 2, 3, 5 and 7-times the input flux period and aperiodic behaviour is verified by measurements of NADH fluorescence in extracts of Saccharomyces cerevisiae in the theoretically predicted range. The stroboscopic transfer function obtained from the aperiodic signal admits period 3, implying chaos according to the Li-Yorke theorem.

Fluorometry↗
PubMed1984

Transitions between oscillatory modes in a glycolytic model system.

A glycolytic model system consisting of the enzymes phosphofructokinase (EC 2.7.1.11) and pyruvate kinase (EC 2.7.1.40) is analyzed when subject to periodic substrate addition. The calculations are performed by using detailed rate laws that have been derived for the enzymes of Escherichia coli. Due to linear relationships between the metabolite concentrations, the numerical solutions can be displayed inside a trapezium, so that the concentrations of four different metabolites are indicated along the trapezium edges. The analysis reveals a rich variety of time patterns, corresponding to different periodic, quasiperiodic, and chaotic attractors. These patterns undergo complex hysteresis loops when bifurcation parameters are slowly changed-for example, by modulating the input amplitude. By using this technique up to four attractors coexisting in phase space are found. The time patterns corresponding to coexisting attractors can be switched into one another by triggering the system with short substrate pulses. Furthermore, conditions exist at which the triggering is autonomous-i.e., self-sustained (intermittent) switchings occur. The time between these switchings can be set externally by the value of the input amplitude. For conditions in which the periods of the oscillations are in the order of minutes, the self-sustained switching-which modulates these oscillations-can be in the order of hours.

Journal Article↗
PubMed1983

Analysis of progress curves. Interaction of pyruvate kinase from Escherichia coli with fructose 1,6-bisphosphate and calcium ions.

The influence of fructose 1,6-bisphosphate and Ca2+ on the kinetics of pyruvate kinase from Escherichia coli K12 was studied (at pH 7.0 and 25 degrees C) by using the pH-stat method for the measurement of the reaction progress as well as initial-rate analysis. The data were analysed on the basis of a concerted model with three conformational states [Markus, Plesser, Boiteux, Hess & Malcovati (1980) Biochem. J. 189, 421-433] by using a novel procedure for a computer-directed treatment of progress curves [Markus & Plesser (1976) Biochem. Soc. Trans. 4, 361-364]. By addition of fructose 1,6-bisphosphate the sigmoid kinetics with respect to phosphoenolpyruvate and Mg2+ is abolished and the activity of the enzyme is described by classical saturation kinetics. This is explained by exclusive binding of fructose 1,6-bisphosphate at an allosteric site of the conformational state that forms the active complex. We observe that Ca2+ is an activator of the enzyme at low Mg2+ and Ca2+ concentrations; otherwise it is an inhibitor. These effects can be understood by assuming that Ca2+ has the same binding properties as Mg2+, although it does not allow a catalytic turnover.

Calcium↗
PubMed1983

Free energy dissipation of the pyruvate kinase reaction has a minimum at cell metabolite concentrations.

The ratio of substrates and products (mass action ratio) for the reaction catalyzed by the enzyme pyruvate kinase is measured under the constraint of constant reaction rate for pyruvate kinase (EC 2.7.1.40) from brewers yeast and Escherichia coli. For both organisms, a maximum of the ratio is found at concentrations comparable to those obtained from cell metabolite measurements. This observation suggests an optimum principle for free energy transduction in the glycolytic reaction pathway, as a maximum of the mass action ratio corresponds to a minimum dissipation of free energy.

Chemical Phenomena↗
PubMed1981

Analysis of progress curves in enzyme kinetics: bias and convergent set in the differential and in the integral method.

Two problems encountered in the analysis of progress curves are examined: 1. Systematic deviations due to errors in the initial solute concentrations make the least-squares method unsuitable. The improvements accomplished by the introduction of a proper weighting matrix are investigated. 2. Non-linear parameter optimization implies a dependence of the optimized parameters on their initial estimates, due to the existence of multiple minima. It is shown that the sensitivity of the optimized parameters on the initial estimates is reduced by fitting the slopes of the progress curves. A subsequent fit of the original progress curve data is recommended for refinement of the parameters.

Enzymes↗
PubMed1980

Analysis of progress curves. Rate law of pyruvate kinase type I from Escherichia coli.

Progress curves of the reaction catalysed by pyruvate kinase from Escherichia coli K12, designed to cover the four-dimensional concentration space of phosphoenolpyruvate, ADP, Mg2+ and ATP in the regulatory region, were recorded with the pH-stat method (pH 7.0 and 25 degrees C). Additional initial-rate measurement were performed to assess specific points. Two methods for the evaluation of progress curves were used: fitting the rate law to the rates obtained from the tangents of the progress curves and fitting the integrated rate law directly to the curves. Two models, both extensions of the concerted model given by Monod, Wyman & Changeux [(1965) J. Mol. Biol. 12, 88--118] with four protomers, could be fitted to the data within the experimental error. Model discrimination in favour of one of these models was possible by proper experimental design. In the selected model one conformational state of the enzyme forms the active complex. The active site of a second conformational state forms abortive complexes with Mg2+, causing strong inhibition at high Mg2+ concentrations. In the absence of ligands, most of the enzyme is in a third state that binds ATP at an allosteric site.

Adenosine Diphosphate↗