Extraction and comparison of bean cytoplasmic and chloroplast ribosomes

1. In isolation of total ribosomes from Phaseolus leaves thehighest yield was obtained with an extraction medium containingHEPES buffer, low Mg²⁺-concentration (1 mM) and SH-group protectingagents. 2. Chloroplast and cytoplasmic ribosomes were obtained in aratio of 1 : 28 after selective isolation. 3. After freezing identical portions of cytoplasmic extractthe amount of ribosomes/ml extract dropped, but later increasedwith freezing time. 4. Ribosomal preparations showed UV-absorption spectra typicalfor RNA. Extinction ratios suggested that preparations werecontaminated with extraneous protein to a certain extent, purityincreasing after selective isolation, especially after freezing. 5. Protein patterns and sedimentation behaviour of cytoplasmicribosomes were identical regardless of addition or omissionof detergent in leaf exeracts. 6. After polyacrylamide gel electrophoresis of ribosomal proteins,29 bands of basic protein were found in cytoplasmic ribosomesand 22 in chloroplast ribosomes. Acidic proteins could not bedetected. Patterns in both extracts were different and highlyreproducible, resembling those recently reported. Tests forfive enzymes, highly active in the leaf tissue, gave negativeresults on the isolated ribosomal proteins. ¹Present address: Max Planck Institut für ExperimentelleMedizin, Arb.-Gr. Biochemie, D-34 Göttingen, Hermann-Rein-Straße3, West Germany. (Received August 10, 1968; )