Partial reconstitution of 60S ribosomal subunits from yeast.
Ribosomal 60S subunits active in polyphenylalanine synthesis can be reconstituted from core particles lacking 20-40% of the total protein. These core particles were obtained by treatment of yeast 60S subunits with dimethylmaleic anhydride, a reagent for protein amino groups. Upon reconstitution a complementary amount of split proteins is incorporated into the ribosomal particles, which have the sedimentation coefficient of the original subunits. Ribosomal protein fractions obtained by extraction with 1.25 M NH4Cl, 4 M LiCl, 7 M LiCl, or 67% acetic acid, are much less efficient in the reconstitution of active subunits from these core particles than the corresponding released fraction prepared with dimethylmaleic anhydride. Attempts to reconstitute active subunits from protein-deficient particles obtained with 1.25 M NH4Cl plus different preparations of ribosomal proteins, including the fraction released with dimethylmaleic anhydride, were unsuccessful. Therefore, under our conditions, of the disassembly procedures assayed only dimethylmaleic anhydride allows partial reconstitution of active 60S subunits.
D000643:Ammonium Chloride; D008299:Maleic Anhydrides; D012270:Ribosomes / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000032:analysis*; D013696:Temperature; D013997:Time Factors
D008299:Maleic Anhydrides; D000643:Ammonium Chloride; C007474:2,3-dimethylmaleic anhydride