Abstract
A method is described for obtaining good yields of purified internal membranes of plastids from greening barley seedlings. The procedure is suitable, without modification, for isolating intact plastids from etiolated seedlings and all early stages of greening. Electron microscopy was used to identify the types of contaminants present in preparations obtained by different methods, and to assess the degree of contamination in purified membranes. Gel electrophoresis of the sodium dodecyl sulphate-solubilised membrane polypeptides established that the proteins pelleted during flotation were a major component of unpurified membrane preparations from etioplasts and plastids greened for 9 hours or less. The polypeptide composition of purified etioplast membranes consisted of 35 bands. At least 15 of these disappeared during subsequent greening. After only 3 hours of illumination, the internal membranes contained almost all of the 43 polypeptides present in mature chloroplast thylakoids. The changes in polypeptide composition from 3–24h were mainly quantitative in nature.
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