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Biomedical subjects

M L Vorbeck

Publications and source records attributed to M L Vorbeck.

At least 19 recordsLinked to original sources
PubMed1990

Effect of aging on the oxidative phosphorylation pathway.

The proposed study was undertaken to investigate the effect of aging on control of the oxidative phosphorylation pathway. Flux control coefficients for adenine nucleotide translocase and cytochrome c oxidase were determined using the procedure of Groen et al. [J. Biol. Chem., 257 (1982) 137-144]. Hepatic mitochondrial fractions from Fischer 344 rats were isolated from control (average age 6.5 months), and aged (average age 27.3 months) groups. No aging-related changes in the extent of control of respiration by the oxidase were obtained, however, differences were observed for the translocase. For the control group of animals, the greatest regulation occurred at 80-85% maximal respiratory rates, and declined at higher rates. For the aged group, a similar flux control coefficient was obtained at 80-85% respiration, but was maintained as respiration increased to maximal rates. It is proposed that changes in the flux control coefficients at maximal respiratory rates are associated with an aging-related decrease in translocase activity. Evaluation of translocase content revealed no significant differences between the two groups supporting the concept that the decreased activity was not due to decreased content. During the course of these experiments, it also became apparent that there was a significant aging-related decrease in the rate of succinate oxidation providing an adequate supply of ADP was present. No significant changes in respiratory rates, or RCR, were evident at suboptimal concentrations of ADP as reported previously from this laboratory [Vorbeck, M.L. et al., Arch. Biochem. Biophys., 214 (1982) 67-79]. Since similar decreases in respiration were obtained upon addition of an uncoupler, the aging-related changes in respiration are attributed to differences at the level of the electron transport system, including its associated reactions. The aging-related differences in respiratory rates, and extent of control of respiration, were both observed under conditions of maximal stimulation of respiration. This suggests an inability of mitochondria from aged animals to respond to the increased demands of oxidation. Basic to these differences may be the lipid-membrane associated changes seen during aging.

Adenosine Diphosphate↗
PubMed1985

Influence of Ca2+ on the plasma membrane potential and electrogenic uptake of glycine by myeloma cells. Involvement of a Ca2+-activated K+ channel.

The involvement of Ca2+-activated K+ channels in the regulation of the plasma membrane potential and electrogenic uptake of glycine in SP 2/0-AG14 lymphocytes was investigated using the potentiometric indicator 3,3'-diethylthiodicarbocyanine iodide. The resting membrane potential was estimated to be -57 +/- 6 mV (n = 4), a value similar to that of normal lymphocytes. The magnitude of the membrane potential and the electrogenic uptake of glycine were dependent on the extracellular K+ concentration, [K+]o, and were significantly enhanced by exogenous calcium. The apparent Vmax of Na+-dependent glycine uptake was doubled in the presence of calcium, whereas the K0.5 was not affected. Ouabain had no influence on the membrane potential under the conditions employed. Additional criteria used to demonstrate the presence of Ca2+-activated K+ channels included the following: (1) addition of EGTA to calcium supplemented cells elicited a rapid depolarization of the membrane potential that was dependent on [K+]o; (2) the calmodulin antagonist, trifluoperazine, depolarized the membrane potential in a dose-dependent and saturable manner with an IC50 of 9.4 microM; and (3) cells treated with the Ca2+-activated K+ channel antagonist, quinine, demonstrated an elevated membrane potential and depressed electrogenic glycine uptake. Results from the present study provide evidence for Ca2+-activated K+ channels in SP 2/0-AG14 lymphocytes, and that their involvement regulates the plasma membrane potential and thereby the electrogenic uptake of Na+-dependent amino acids.

Animals↗
PubMed1984

An ELISA for screening hybridoma cultures for monoclonal antibodies against a detergent solubilized integral membrane protein.

A method is described for the binding of a detergent solubilized integral membrane protein to polystyrene immunoassay plates. Addition of Bouin's fluid, a histochemical fixative, to wells of plates containing the detergent solubilized antigen, followed by low speed centrifugation, is sufficient to promote binding of antigen in the presence of Triton X-100 concentrations as high as 1.75%. The binding of antigen is rapid and the entire binding procedure, including removal of fixative and washing of the plates, can be accomplished in less than 15 min. Immunological specificity of the bound antigen is retained. This method has been used to effectively screen hybridoma cultures for specific antibodies.

Acetates↗
PubMed1983

Ca2+ release from energetically coupled tumor mitochondria.

A Na+/Ca2+ exchange activity for Ca2+ efflux has been identified in isolated Ehrlich ascites tumor mitochondria. Further, under conditions favoring cycling of Ca2+ across the mitochondrial inner membrane, extramitochondrial [Ca2+] also was shown to be Na+-dependent. The Na+/Ca2+ exchange showed sigmoidal kinetics with a mean (+/-SD) [Na+] required for half maximal stimulation of Ca2+ efflux of 8.4 +/- 3.8 mM and a Hill coefficient of 1.6. Na+/Ca2+ exchange was very sensitive to inhibition by the Ca2+ antagonist diltiazem (56% inhibition at 7.5 nmoles X mg protein-1) whereas a number of other compounds, including verapamil, nupercaine, and trifluoperazine were less effective in inhibiting Ca2+ efflux. These data demonstrate for the first time the presence of a pathway in tumor mitochondria for unidirectional Ca2+ efflux induced by Na+, and provide a mechanism for regulation of tumor intra- and extramitochondrial [Ca2+]. Results of the present study support the need for further study of intracellular Na+ and its role in regulation of Ca2+ homeostasis in tumor cells.

Animals↗
PubMed1982

Ca2+-dependent depolarization of energized mitochondrial membrane potential by chlortetracycline (aureomycin).

The addition of chlortetracycline (CTC) to succinate-energized rat liver mitochondria resulted in depolarization of the membrane potential and decreased respiratory control. CTC inhibited both processes at concentrations that were half maximally effective at approximately 13 and 16 microM, respectively. These inhibitory effects were prevented by either the Ca2+ chelator, ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, or the inhibitor of mitochondrial Ca2+ influx, ruthenium red. These findings are consistent with the formation of a membrane associated calcium-CTC complex and suggest that CTC can alter mitochondrial energy metabolism during transmembrane Ca2+ cycling.

Adenosine Diphosphate↗
PubMed1980

Computerized 3-dimensional analysis of mitochondrial structure.

A computer program has been written to calculate surface area and volume of subcellular organelles. Sophisticated and extremely accurate point sampling routines and volume computational techniques have been developed. This program has been used to study mitochondrial 3-dimensional structure when stereological estimates have proven inadequate.

Animals↗
PubMed1975

Membrane phospholipid metabolism in the isoproterenol-induced cardiomyopathy of the rat.

Within 60 minutes after treatment with low doses of IPR (5 mg/kg), relative specific activity of cardiolipin and sphingomyelin significantly increased in the heart. Other phospholipids of the heart were not significantly changed. Relative specific activity of cardiolipin increased much later in liver and salivary gland. Activity of CTP:PA cytidyl transferase increases 40% at 1 hour following treatment with IPR. No significant change was observed in the respiratory control ratio of heart mitochondria for 2 hours following injection with IPR; however, a significant decrease was observed 24 hours after IPR.

Animals↗