Incorporation of Cystine by Yeast Cells of Histoplasma capsulatum.

This report deals with the incorporation of cystine into macromolecules by yeast cells of Histoplasma capsulatum. The results show that at saturating concentrations of cystine in a rich medium, total uptake of the cystine occurs in 10 hr, whereas the amount of label in the cold trichloroacetic acid-soluble material reaches a maximum at 3 to 4 hr and remains at this value. The amount of label in the cold acid-insoluble material accumulates linearly for 4 to 5 hr and reaches a plateau at 7 to 9 hr. A chemical fractionation of labeled cells shows that 25% of the cystine taken up remains as low-molecular-weight components, of which cystine comprises 60 to 75%. Approximately 30% of the total label incorporated is ethanol-soluble, and polyacrylamide gel electrophoresis of this material shows a rather uniform incorporation of the amino acid into many proteins. The combined hot KOH fractions account for 40% of the total label incorporated. The amount of hot KOH-insoluble material almost doubles in a 10-hr pulse, whereas there is an increase in hot KOH-soluble material. Hence, the greatest amount of label from cells pulsed with (14)C-cystine is recovered from cell wall material.