Sub-thylakoid fractions containing ribosomes.
A sub-membrane fraction which contains a large portion of any thylakoid-bound ribosomes can be obtained when thylakoids are treated with the detergents Nonidet P-40, or Triton X-100. These 'pseudopolysome' fractions contain 50% of thylakoid-bound ribosomes, but less than 0.5% of thylakoid chlorophyll. Triton and Nonidet psuedopolysomes contain about 10%, and 3% of thylakoid protein, respectively. Pseudopolysomes, prepared from thylakoids with low levels of ribosomes, contain about the same proportion of thylakoid protein but proportionately less ribosomes. Pseudopolysomes contain thylakoid polypeptides in addition to chlorophyll, but lack a major membrane polypeptide of Mr 50 000. Pseudopolysome chlorophyll, and RNA band at the same buoyant density. However, they band at different densities after pseudopolysomes are treated with trypsin (a procedure which strips thylakoids of ribosomes). Pseudopolysome fractions from thylakoids with low levels of ribosomes have a lower density than the corresponding fractions from thylakoids with high levels of ribosomes. Ribosomes are released from thylakoids, and pseudopolysomes by the same treatments. Subunits are released with KCl and puromycin. Polysomes are released with trypsin. It was concluded the pseudopolysomes consist of ribosomes and a membrane fragment containing the sites to which ribosomes are bound.
D002696:Chlamydomonas / Q000032:analysis; D002734:Chlorophyll / Q000032:analysis; D002736:Chloroplasts / Q000032:analysis*; D008566:Membranes / Q000032:analysis; D010455:Peptides / Q000032:analysis; D011092:Polyethylene Glycols; D011132:Polyribosomes / Q000032:analysis; D011189:Potassium Chloride; D011691:Puromycin; D012335:RNA, Ribosomal / Q000032:analysis; D012270:Ribosomes / Q000032:analysis*; D014357:Trypsin
D010455:Peptides; D012335:RNA, Ribosomal; D002734:Chlorophyll; C024062:nonidet; D011092:Polyethylene Glycols; D011691:Puromycin; D011189:Potassium Chloride; D014357:Trypsin