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G Jacobasch

Publications and source records attributed to G Jacobasch.

At least 55 records · Page 3Linked to original sources
PubMed1989

Mathematical modelling of metabolic pathways affected by an enzyme deficiency. Energy and redox metabolism of glucose-6-phosphate-dehydrogenase-deficient erythrocytes.

The effects of various forms of glucose-6-phosphate dehydrogenase deficiency on erythrocyte metabolism have been studied on the basis of a complex mathematical model which comprises the main pathways of this cell: glycolysis, pentose pathway, reactions of the glutathione and adenine nucleotide metabolism. The calculated flux rates through the oxidative pentose pathway with and without methylene blue are in good accord with experimental results. The degree of deficiency as predicted by the model on the basis of calculated upper oxidative load boundaries, as well as of maximal methylene blue stimulation, correlates with the individual clinical manifestation of the metabolic disease. Therefore, the model allows one to judge the degree of metabolic disorder in the presence of glucose-6-phosphate dehydrogenase enzymopathies if the kinetic properties of the defect enzyme are known. Experimentally accessible parameters for an assessment of the oxidative load capacity of cells in vivo are proposed. It is pointed out that the threshold of tolerance as to energetic load is drastically reduced in the case of severe glucose-6-phosphate dehydrogenase deficiency.

Catalysis↗
PubMed1989

[Clinico-biochemical characteristics of patients with congenital glucose-6-phosphate dehydrogenase deficiency in erythrocytes].

The authors submit a clinical and biochemical investigation of five families with a congenital glucose-6-phosphate dehydrogenase red cell deficiency. Congenital nonspherocytic haemolytic anaemia was detected only in one hemizygous carrier, in four there was latent haemolysis with intermittent haemolytic crises and hyperbilirubinaemia. In 14 relatives carriership was revealed without clinical manifestations. The authors give a biochemical characteristic of kinetic properties of enzymatic variants according to WHO instructions (enzyme activity, thermal stability, pH stability and other kinetic constant) and emphasize the need to improve the diagnosis.

Adolescent↗
PubMed1988

A kinetic model of phosphofructokinase from Plasmodium berghei. Influence of ATP and fructose-6-phosphate.

Phosphofructokinase (PFK) from the malarial parasite Plasmodium berghei shows the following kinetic features: the more the pH is decreased, the more the enzyme is inhibited by ATP; in contrast to PFK from erythrocytes, this inhibition is less potent by two orders of magnitude; as in the red cell, fructose-6-phosphate (F6P) is a positive effector. Kinetic modelling of PFK from P. berghei has been performed by taking the pH-dependence of activity into regard, implicitly by the estimation of pH-dependent kinetic parameters for the inhibition by ATP and the activation by F6P and explicitly by the assumption of protonation-steps involved in allosteric regulation. By means of a novel procedure of model discrimination [D. Buckwitz and H.-G. Holzhütter: A new method to discriminate between enzyme-kinetic models. In: Application of Computational Methods in Medicine (Györi, I., ed.), Akademai, Budapest, in press] we have selected among several kinetic models the best rate equation which provides an adequate quantitative description of the kinetic behaviour of the enzyme in the relevant ranges of substrate concentrations and pH (5.8-7.6). It thus becomes clear how the highly increased glycolytic flux in malaria-infected cells could be affected through PFK.

Adenosine Triphosphate↗
PubMed1988

Interrelations between glycolysis and the hexose monophosphate shunt in erythrocytes as studied on the basis of a mathematical model.

A mathematical model is presented which comprises the reactions of glycolysis, the hexose monophosphate shunt (HMS) and the glutathione system in erythrocytes. The model is used to calculate stationary and time-dependent metabolic states of the cell in vitro and in vivo. The model properly accounts for the following metabolic features observed in vitro: (a) stimulation of the oxidative pentose pathway after addition of pyruvate due to a NADP-dependent lactate dehydrogenase as coupling enzyme between glycolysis and the oxidative pentose pathway, (b) relative share of the oxidative pentose pathway in the total consumption of glucose amounting to approximately 10% in the normal case and to approximately 90% under conditions of oxidative stress excreted by methylene blue. From the application of the model to in vivo conditions it is predicted that (c) under normal conditions glycolysis and the HMS are independently regulated by the energetic and oxidative load, respectively, (d) under conditions of enhanced energetic or oxidative load both glycolysis and the HMS are mainly controlled by the hexokinase; in this situation the highest possible values of the energetic and oxidative load which are compatible with cell integrity are strongly coupled and considerably restricted in comparison with the normal case, (e) the stationary states possess bifurcation points at high and low values of the energetic load.

Energy Metabolism↗
PubMed1987

Membrane elastic shear modulus of red blood cells with glucose-6-phosphate dehydrogenase and pyruvatekinase enzymopathies.

The membrane elastic shear modulus mu determined by a micropipette technique was found to be elevated by 25% to 200% for red blood cells (RBC) from 7 patients with glucose-6-phosphate dehydrogenase (G-6-PD) and 9 patients with pyruvate-kinase (PK) enzymopathies above the mean value for normal controls. All patients exhibit chronic nonspherocytic hemolytic anemias. A negative linear correlation (r = -0.72) between mu and the number of reticulocytes was obtained in G-6-PD deficiencies. In contrast, a positive correlation (r = 0.88) was found for red blood cells of patients with PK deficiency. A mechanically induced swelling of RBC was observed for some deficient cells. The results are discussed in the framework of structural alterations of the RBC membrane due to the disturbed pentose-phosphate pathway and the diminished ATP supply through glycolysis, respectively.

Elasticity↗
PubMed1987

Heterogeneity of glucose-6-phosphate dehydrogenase enzymopathies in the GDR.

G6PD variants of 13 patients from 12 German families with different clinical symptoms have been characterized kinetically. Vmax G6PD was nearly zero in red blood cells of all carriers. Therefore G6PD variants were isolated from leucocytes, which proved to be a suitable source for analysis of instable G6PD variants. The testing program included KmG6P, KmNADP, Ki values of NADPH, ATP and 2,3 P2G, rate of utilization of dG6P, Gal6P, dNADP, NAD, and pH dependence. From the results obtained one can conclude that all analyzed G6PD variants represent individual mutations. The degree of metabolic dysregulation can be explained by the different kinetic and physico-chemical properties of these G6PD variants.

Erythrocytes↗
PubMed1987

Human erythrocyte pyruvate kinase (L'/R-PK): production and characterization of a monoclonal antibody.

Two preparations of L'/R-type pyruvate kinase from human erythrocytes characterized by SDS-PAGE were used for immunization of BALB/c mice. Their spleen cells were fused with mouse myeloma cells by polyethylene glycol according to standard techniques. Supernatants of hybridomas resulting from two separate experiments were assayed by ELISA and further characterized by immunoblotting. Using those monoclonal antibodies reacting with L'/R-PK in immunoblotting, a major band of 62 KDa MW was recognized in both preparations employed for immunization. Additionally, four smaller bands were detected. Furthermore, the monoclonal antibodies also detected a single band of 62 KDa MW in a highly purified L-PK prepared from human liver. In contrast, they showed no reaction with muscle and brain tissues containing M1-type PK. However, they reacted strongly with a single band of approximately 62 KDa in liver and kidney homogenates which is in line with immunocytochemical studies showing immunoreactive material in hepatocytes and proximal tubules of the kidney.

Antibodies, Monoclonal↗
PubMed1986

Kinetic model of glucose-6-phosphate dehydrogenase from red blood cells. Parameter estimation from progress curves and simulation of regulatory properties.

A kinetic model of human and mouse glucose-6-phosphate dehydrogenase is presented which takes into account the substrates and all inhibitors of significant importance in the red cell. The parameter values were estimated by analysis of progress curves. The applicability of a new method based on non-linear regression to complex enzyme kinetics was proved. The in vivo-regulation of glucose-6-phosphate dehydrogenase is examined by determining elasticity coefficients and by using simple simulation experiments. The model is convenient to describe the behaviour of enzyme activity under physiological conditions.

Adenosine Triphosphate↗
PubMed1985

Mathematical modelling of metabolic pathways affected by an enzyme deficiency. A mathematical model of glycolysis in normal and pyruvate-kinase-deficient red blood cells.

A mathematical model of glycolysis in human erythrocytes is proposed to study the influence of a pyruvate kinase deficiency on the energy metabolism. The model takes into account the main regulatory properties of the non-equilibrium enzymes and the magnesium-complex formation by the adenine nucleotides and by 2,3-bisphosphoglycerate. In the normal case (no enzyme defect) the calculated flux rates and metabolite concentrations are in a good agreement with experimental data. It is shown that a severe pyruvate kinase deficiency manifested in a tenfold diminished activity of that enzyme leads to a remarkable decrease of the glycolytic flux and the ATP concentration of about 50% of the normal values. On the other hand a lowering of the pyruvate kinase activity to half of the normal value, characteristic for the heterozygotes, gives no significant alterations of the metabolite concentrations and the flux rates compared with the normal case which is in accordance with the lack of clinical symptoms for a metabolic disease of these probands. For three patients with known alterations of their pyruvate kinase mutants the calculated metabolite concentrations and the control characteristics permit estimation of the degree of disorder of the glycolytic pathway. The resulting classification corresponds well to other independent experimental and clinical findings. In particular, the calculation demonstrates that there is no simple correlation between the lowered enzyme activity and the reduced flux rate through the affected pathway.

Adenosine Triphosphate↗
PubMed1984

[Proteins and phospholipids of thrombocytes and erythrocytes in glucose-6-phosphate dehydrogenase deficient patients].

Proteins and phospholipids of platelets and red cell membranes of glucose-6-phosphate-dehydrogenase-deficient patients with chronic hemolytic disease were investigated. The enzyme deficiency is not connected with alteration in the protein and phospholipid pattern of platelets. However, marked differences in the diamide-induced protein polymerization between normal and deficient platelets were found. Whereas normal platelets show only a weak polymerization which is reversible during diamide incubation of the cells, the deficient platelets demonstrate a considerable and long lasting protein polymerization. The results point to a direct connection between the thiol-disulfide status of platelets and intracellular level of glutathione, which is able to repair oxidative damages. Two of the six patients showed already protein polymers in their erythrocyte membrane without addition of any SH-oxidizing agent. The phospholipid pattern of the deficient red-cell membranes were comparable to the controls.

Adolescent↗
PubMed1984

Molecular and cellular regulation of pyruvate kinase in red blood cells.

The main sources of the heterogeneity of pyruvate kinase (PK) isoenzymes are discussed. The regulative role of the L-type PK for the energy metabolism of a tissue without glyconeogenesis has been examined in erythrocytes. For that purpose a kinetic model of the PK has been established taking into account the substrates and the effectors ATP and fructose-1,6.biphosphate. This PK model was incorporated into a complex mathematical model of glycolysis and the concentrations of metabolites and the glycolytic flux were calculated for normal PK activity and for a PK activity ten times lower than normal. From calculations it is concluded that under the conditions of PK deficiency the energy metabolism cannot be stabilized. This corresponds to the experimental finding that the filtration constant of PK deficient red blood cells increases with declining survival time.

Erythrocytes↗
PubMed1983

Mechanical membrane properties of human red blood cells and their change due to metabolic disturbance.

Blood of patients suffering from non-spherocytic chronic haemolytic anemia due to individual mutations of glucose-6-phosphate-dehydrogenase (G-6-PD) and pyruvate kinase (PK) was investigated by biochemical and rheological methods. Both enzymopathies are characterized by a strong increase (by the factor 2 to 4) in the overall RBC-rigidity and the elastic membrane shear modulus of the RBC. There was a positive correlation between the changed elastic shear modulus and the clinical picture in the case of PK deficiency and, in contrast, a negative correlation for patients with G-6-PD enzymopathies. The ability of RBC to change their shape or the static deformability is determined by the excess surface regarding to the enclosed volume, by the rheological properties of the hemoglobin content and by the membrane extension and bending moduli. The dynamic deformability is characterized by the time constant for rapid elastic and plastic deformations and becomes important for entrance and discharge processes in the microcirculation. These rheological relevant properties are subjected to metabolic control. Although for both enzymopathies the mechanism of hemolysis is not understood in detail, it has to be assumed that PK deficient RBC are mostly phagocytized and on the other hand G-6-PD deficient cells are destroyed to a higher extent by intravasal hemolysis. The aim of this study was to investigate whether or not expected changes in the RBC membrane structure due to the decreased ATP production by PK enzymopathies and diminished NADPH production by G-6-PD deficiencies result in abnormal mechanical membrane properties. The answer should give a better understanding of the premature RBC destruction.

Anemia, Hemolytic↗
PubMed1983

The energy metabolism of pyruvate kinase deficient red blood cells.

The metabolic consequences of PK enzymopathies on the energy metabolism have been studied by means of mathematical modelling. The proposed steady state model of glycolysis takes into account the main regulatory properties of the non-equilibrium enzymes and the Mg-complex formation by the adenine nucleotides and 2,3-bisphosphoglycerate. The analysis of three distinct individual cases of PK enzymopathies has demonstrated that mathematical modelling may lead to a deeper understanding and a quantitative estimate of the metabolic disregulation.

Energy Metabolism↗