Total Reconstitution of Functionally Active 50S Ribosomal Subunits from Escherichia coli
- 1 December 1974
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 71 (12) , 4713-4717
- https://doi.org/10.1073/pnas.71.12.4713
Abstract
Total reconstitution of 50S subunits from E. coli was achieved by a two-step incubation procedure. In the first step, 23S RNA, 5S RNA, and the total proteins from 50S subunits were incubated for 20 min at 40 degrees in the presence of 4 mM Mg(++) and 400 mM NH(4)Cl. In the second step, the Mg(++) concentration was raised to 20 mM and the incubation was performed for 90 min at 50 degrees . No requirement for 30S subunits or other components (e.g., polyamine) was found. The reconstituted particle has the same sedimentation coefficient as the native 50S subunit and is highly active in protein synthesis with natural (R17 RNA) and artificial [poly(U)] messengers as well as in tests for peptidyltransferase (fragment assay) and for binding of antibiotics (chloramphenicol).Keywords
This publication has 23 references indexed in Scilit:
- The structural basis for functional inactivity of reconstituted 50-S ribosomal subunits of Escherichia coliBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1973
- Identification of the Chloramphenicol-Binding Protein in Escherichia coli Ribosomes by Partial ReconstitutionProceedings of the National Academy of Sciences, 1973
- A Protein Involved in the Peptidyltransferase Activity of Escherichia coli RibosomesProceedings of the National Academy of Sciences, 1973
- Action of methanol on the association of ribosomal subunits and its effect on the GTPase activity of elongation factor GBiochemical and Biophysical Research Communications, 1973
- Association of a ribonuclease with the 50-S ribosomal subunit of Escherichia coli MRE 600Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1973
- Ribosomal proteins: XLIII. In vivo assembly of Escherichia coli ribosomal proteinsJournal of Molecular Biology, 1973
- Physical Reconstitution of 23 S RNA – 50 S Protein Complexes from Escherichia coliCanadian Journal of Biochemistry, 1973
- Tables for estimating sedimentation through linear concentration gradients of sucrose solutionAnalytical Biochemistry, 1967
- In vitro recovery of ribosomes and of synthetic activity from synthetically inactive ribosomal subunitsJournal of Molecular Biology, 1966
- Reconstitution of functionally active ribosomes from inactive subparticles and proteins.Proceedings of the National Academy of Sciences, 1966