Analysis of the energy metabolism after incubation of Saccharomyces cerevisiae with sulfite or nitrite

After addition of 5 mM sulfite or nitrite to glucose-metabolizing cells of Saccharomyces cerevisiae a rapid decrease of the ATP content and an inversely proportional increase in the level of inorganic phosphate was observed. The concentration of ADP shows only small and transient changes. Cells of the yeast mutant pet 936, lacking mitochondrial F₁ATPase, after addition of 5 mM sulfite or nitrite exhibit changes in ATP, ADP and inorganic phosphate very similar to those observed in wild type cells. They key enzyme of glucose degradation, glyceraldehyde-3-phosphate dehydrogenase was previously shown to be the most sulfiteor nitrite-sensitive enzyme of the glycolytic pathway. This enzyme shows the same sensitivity to sulfite or nitrite in cells of the mutant pet 936 as in wild type cells. It is concluded that the effects of sulfite or nitrite on ATP, ADP and inorganic phosphate are the result of inhibition of glyceraldehyde-3-phosphate dehydrogenase and not of inhibition of phosphorylation processes in the mitochondria. Levels of GTP, UTP and CTP show parallel changes to ATP. This is explained by the presence of very active nucleoside monophosphate kinases which cause a rapid exchange between the nucleoside phosphates. The effects of the sudden inhibition of glucose degradation by sulfite or nitrite on levels of ATP, ADP and inorganic phosphate are discussed in terms of the theory of Lynen (1942) on compensating phosphorylation and dephosphorylation in steady state glucose metabolizing yeast.Abbreviations ATP adenosine triphosphate - ADP adenosine diphosphate - AMP adenosine monophosphate - P_i inorganic orthophosphate Dedicated to Prof. Dr. Hans Grisebach on the occasion of his sixtieth birthday     

Zur Frage einer intraventrikulären Neurosekretion

Zusammenfassung In der vorderen Wand des IV. Ventrikels und des Canalis centralis gegenüber der Area postrema liegen beim Kaninchen Nervenzellen, deren Fortsätze intraventrikulär mit 6 großen Protoplasmakugeln endigen, die mit Mitochondrien angefüllt sind. Das Perikaryon ist reich an Ergastoplasma, alle Teile der Zelle enthalten leere, 600 bis 1500 Å messende, und ein dichtes Granulum enthaltende, 650–1000 Å große Bläschen. Vermutlich werden Stoffe in den Liquor cerebrospinalis abgegeben. Dem Plasmalemm der Nervenzellen liegen Synapsen an.

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Summary In the anterior wall of the IVth ventricle, opposite to the area postrema, nerve cells are found. Their processes end with spheroid swellings extending into the ventricle. These bulbs contain a large number of mitochondria, while ergastoplasm is seen in the perikaryon. The cytoplasm of the whole cell shows vesicles (600–1500 Å) and dense-core vesicles (650–1000 Å) which might be extruded into the liquor cerebrospinalis. The plasmalemma of the perikaryon is in contact with synapses.

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Die Untersuchung wurde mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Antioxidant Activity of 5-Hydroxytryptophan, 5-Hydroxyindole, and Dopa Against Microsomal Lipid Peroxidation and Its Dependence on Vitamin E

The antioxidant capacity of 5-hydroxy-tryptophan. 5-hydroxy-indole. and DOPA (3,4-dihydroxy-phenyI-alanine) was tested in the Fe-induced lipid peroxidation of liver microsomes of normal- and vitamin E-deficient rats, using ascorbate as a reductant. Lipid peroxidation was monitored as low-level chemilu-minescence, indicative of generation of electronically-excited states arising from the recombination of secondary lipid peroxyl radicals.     

Über die elektrische Kapazität der Zellmembran und die Leitfähigkeit des Zytoplasmas von Ehrlich-Aszitestumorzellen

Zusammenfassung In der vorliegenden Arbeit wird die elektrische Leitfähigkeit und die Membrankapazität von Ehrlich-Aszitestumorzellen gemessen. Die Membrankapazität beträgt 1,7±0,3 Farad/cm². Die spezifische Leitfähigkeit des Zytoplasmas beträgt 0,013 [ ^–1 cm^–1]±12%. Der spezielle Verlauf der Dispersion der Dielektrizitätskonstanten und der Leitfähigkeit der Zellsuspension zeigt, daß ein sehr breites Spektrum von Relaxationszeiten vorliegt, das nicht durch die Größenverteilung der Zellen allein erklärt werden kann. Die Spektralverteilung der Relaxationszeiten hat die FormH(T)=const. Die spezifische Leitfähigkeit des Protoplasmas kann in erster Näherung durch die elektrische Beweglichkeit der Elektrolytionen in einer etwa 15%igen Proteinlösung erklärt werden.Für die Mitarbeit bei den Versuchen möchte ich Frau H.Valetas meinen Dank aussprechen.Herrn Prof. Dr. Dr. h. c. Dr. h. c.Boris Rajewsky zum 70. Geburtstag gewidmet.     

Entwicklungsgeschichtliche untersuchungen an einer Blattmutante vonHyoscyamus niger L.

Summary The leaf shape of the mutantfiliformis (fil) ofHyoscyamus niger L. is strongly modified by external factors (like nutrition and light) as well as by the height of insertion. The name filiformis refers to thread-like leaves which always occur in the inflorencence; they may also be formed in the vegetative region, especially under short day conditions. Other leaves may have a small rhombic blade or a larger blade with irregular edges and deep incisions. Even pinnate leaves have been found. In contrast to the leaves of normalHyoscyamus, all mutant leaves (hypsophylls included) have a stalk-like basal portion that seems to be homologous to the basal part of the normal blade. This mutant is caused by one recessive factor which is linked neither toann nor topall.the submarginal initials of the normalHyoscyamus blade were always found dividing according to the periclinal-anticlinal type, while in the mutant the activity of the submarginal initials frequently resulted in a primarily biseriate mesophyll (so-called double-edged segmentation).This is apparently the first time that gene control of the mode of submarginal blade growth has been observed. Further differences between mutant and normalHyoscyamus concern the venation, the lengths of palisade cells and of stomata guard cells, the frequency of stomata per mm², and the thickness of the blade.

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Mit 7 Textabbildungen     

The leaf peroxisomal form (MFP IV) of multifunctional protein functioning in fatty-acid β-oxidation

We have purified for the first time from green leaves a multifunctional protein (MFP) involved in fatty acid -oxidation. The protein, designated MFP IV, was extracted from green leaves of three-week-old cucumber (Cucumis sativus L.) plants. Chromatography on cation exchanger, separation on hydroxylapatite, and fast-protein liquid chromatography on Phenylsuperose led to a more than 7000-fold purification and to the isolation of an apparently homogeneous 80-kDa monomeric protein. This protein is immunologically related to the glyoxysomal MFP II, as evidenced by immunodecoration with antiserum raised against MFP II. Comparison of molecular masses of all MFPs presently known revealed that the MFP prepared from green leaves (MFP IV) is distinct from MFP II (76.5 kDa) and MFP I (74 kDa) from dark-grown cotyledons. By including other properties in this comparison, we demonstrated that MFP IV can also be distinguished from the glyoxysomal MFP III (81 kDa) and the bacterially expressed MFP-a (80 kDa). Moreover, MFP IV is a constituent of leaf peroxisomes and contains the activities of 2-enoyl-CoA hydratase (EC 4.2.1.17),l-3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) and 3-hydroxyacyl-CoA epimerase.Abbreviation MFP multifunctional protein This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie.     

Cytochrome b 558 and hydrogen peroxide production in small intensely fluorescent cells of sympathetic ganglia

 Small intensely fluorescent (SIF) cells are paraganglionic cells derived from sympathicoblasts which may serve as interneurons, endo-/paracrine cells or arterial chemoreceptors within sympathetic ganglia. Like paraganglionic cells of other locations, e.g., carotid body glomus cells, they are responsive to hypoxia. Recent studies on glomus cells and other hypoxia-sensing cells suggested the involvement of a b ₅₅₈ -type cytochrome and intracellular generation of H₂O₂ in the process of oxygen sensing. In the present study, we demonstrate the occurrence of the small subunit of cytochrome b ₅₅₈ , p22phox, in SIF cells of guinea-pig sympathetic ganglia by immunohistochemistry using two different antisera. H₂O₂ production was monitored in explanted intact superior cervical ganglia of 2-day-old rats by confocal laser scanning analysis of rhodamine 123 fluorescence generated due to oxidation of dihydrorhodamine 123 by H₂O₂. Using this technique, SIF cell clusters appeared as sites of highest H₂O₂ production within the ganglia. Thus, SIF cells exhibit two key features of an oxidase system generating reactive oxygen species. This may be involved in the proposed chain of events in oxygen sensing, but alternative cellular functions of this system have also to be considered. Accepted: 19 September 1996