Stabilization,solubilization, partial purification and some properties of cytochrome P-450 present in CaCl2-precipitated human placental microsomes

Two forms of a carbon monoxide-binding cytochrome were found to exist in CaCl₂-precipitated microsomal subfractions of human placental homogenates at term. These exhibited absorption maxima at wavelengths of 450 nm and 421–432 nm. Conversion of P-450 to the degradation product could be prevented by resuspension of the microsomes in buffered solutions containing 20% glycerol and 10^?4M dithiothreitol. Solubilization of such resuspended microsomes with sodium cholate could be effected with minimal degradation. The solubilized pigment then could be partially purified by differential fractionation with ammonium sulfate. The partially purified cytochromes did not appear to bind desmethylimpramine but did bind aniline and nicotinamide to yield atypical type II difference spectra with maxima at 435 nm and minima at 416 nm. A type I difference spectrum could be elicited with androstenedione and a spectral dissociation constant (K_S) of 4.7×10^?8M was obtained. Androstenedione also appeared to effectively prevent the binding of carbon monoxide to the cytochrome.     

The Biological Clock in Chlamydomonas reinhardi

SYNOPSIS. Chlamydomonas reinhardi has a biological clock regulating phototaxis in dividing and non-dividing cultures; it also can exert some control on growth of continuous cultures. The period length is ∼ 24 hr; it is temperature-compensated and not dependent on the average growth rate. The rhythm can be entrained or phased by light-dark conditions. In dividing cultures a periodic fluctuation in cell number and total protein persists in continuous light.     

Über die Ursachen der Osmotoleranz bei Hefen

Zusammenfassung Mit Untersuchungen über die Abhängigkeit des Zellvolumens vom osmotischen Wert und über die Konzentration der Zellinhaltsstoffe wird versucht, zur Kenntnis der Ursachen der Osmotoleranz bei Hefen beizutragen. In konzentrierten Lösungen verringern alle untersuchten Hefen ihr Volumen, die osmotoleranten Hefen aber weniger als die nichtosmotoleranten. Der Beginn einer meßbaren Volumenverringerung tritt bei osmotoleranten Hefen bei höheren Konzentrationen ein als bei nichtosmotoleranten. Eine Abhängigkeit der Zellgröße vom osmotischen Wert kann nur in einem bestimmten Konzentrationsbereich (zwischen 0,7–2% und 10% NaCl) festgestellt werden. Darunter fehlt eine osmotische Reaktion, und darüber bleibt die Zellgröße gleich. In Nährlösungen mit erhöhtem osmotischen Wert vermehren sich Zellen beider Typen in ihrer normalen Größe. Der osmotische Wert ihres Zellinhaltes ist annähernd gleich groß. Eine veränderte Struktur des Zellplasmas bzw. ein dadurch bedingter höherer Gehalt an fest gebundenem Hydratationswasser wird als Ursache der Osmotoleranz bei Hefen angesehen.

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About the causes of osmotolerance in yeasts

Summary The question, if the yeast cell volume depends on the osmotic pressure and on the solutes of the cell, has been investigated. In concentrated solutions all yeasts investigated reduced their volume. A smaller reduction was found in osmotolerant yeasts. Osmotolerant yeasts begin to reduce their volume at higher osmotic pressures than non-osmotolerant ones. In concentrated nutrient media the cells of osmotolerant and non-osmotolerant yeasts grow in their normal size. The cell size depends only in the range between 0.7 to 2% and 10% NaCl approximately on the osmotic pressure of the surrounding medium. Beneath 2% there is no osmotic reaction and above 10% the cell size remains unchanged. The osmotic value in the cells of both yeast types is about the same. Causes of osmotolerance in yeasts may be a modified structure of cytoplasm and an increased content of bound water.

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(Prof. Dr. S. Windisch)