b-type cytochromes in plasma membranes of Phaseolus vulgaris hypocotyls, Arabidopsis thaliana leaves, and Zea mays roots.

The plasma membrane of higher plants contains more than one kind of b-type cytochromes. One of these has a high redox potential and can be fully reduced by ascorbate. This component, the cytochrome b561 (cyt b561), has its characteristic alpha-band absorbance close to 561 nm wavelength at room temperature. Cyt b561 was first isolated from etiolated bean hook plasma membranes by two consecutive anion exchange chromatography steps. During the first step performed at pH 8, cyt b561 did not bind to the anion exchange column, but other b-type cytochromes did. In the second step performed at pH 9.9, cyt b561 was bound to the column and was eluted from the column at an ionic strength of about 100 mM KCl. However, when the same protocol was applied to the solubilized plasma membrane proteins from Arabidopsis thaliana leaves and maize roots, the ascorbate-reducible cyt b561 bound already to the first anion exchange column at pH 8 and was eluted also at an ionic strength of about 100 mM KCl. Other b-type cytochromes than the ascorbate-reducible cyt b561 from the plasma membranes of Arabidopsis leaves and maize roots showed similar chromatographic characteristics to that of bean hypocotyls. These results demonstrate particular differences in the chromatographic behavior of cyt b561 from different sources.