Méthode d'estimation de la concentration des cytochromes dans les cellules entières de levure

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Abstract
A method for the estimation of cytochrome concentrations in whole yeast cells was worked out with strains of Saccharomyces cerevisiae obtained in the laboratory. Differences of cytochrome c content were due to mutations affecting the nature or the quantity of synthetized cytochrome c. Respiratory deficient mutants (without cytochromes a+a3 and b) were also used. Spectra were measured directly on yeast pastes (up to 3 mm thick) with a Cary 15 recording spectrophotometer provided with a Xenon lamp.Absorption differences between the maxima of α and β peaks of cytochrome c in relation to two base lines were measured. The mean value so obtained was in accordance with Lambert's law when the thickness of the sample of the same yeast paste was changed. For a constant optical path, Beer's law has been verified with the strains of different cytochrome c contents due to genotypical differences. There is a linear relation between quantities of cytochrome c extracted and absorptions measured on whole cells pastes.A device for liquid nitrogen temperature recordings is described and the extension of the method to low temperature measurements, and to the estimation of cytochromes a+a3 concentration is discussed.

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