Open Access50‐S Subunit from Escherichia coli Ribosomes
Author(s) -
WYSTUP Gabriele,
TERAOKA Hiroshi,
SCHULZE Hartmut,
HAMPL Hartmut,
NIERHAUS Knud H.
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb02038.x
Subject(s) - ribosome , protein subunit , escherichia coli , size exclusion chromatography , sephadex , ribosomal protein , chemistry , eukaryotic large ribosomal subunit , gel electrophoresis , biochemistry , ion chromatography , chromatography , ribosomal rna , polyacrylamide gel electrophoresis , urea , enzyme , rna , gene
A method is described for the isolation of highly purified proteins from the 50‐S subunit of Escherichia coli ribosomes. All the proteins from the large subunit could be isolated with the exception of L14, L26, L31 and L34. The isolated proteins are functionally active in reconstituted particles. The method consists of successive NH 4 Cl/EtOH and LiCl washing steps, which split off distinct groups of proteins from the ribosome. The protein groups are further separated by a combination of gel filtration (Sephadex G‐100) and ion‐exchange chromatography (carboxymethylcellulose) in the presence of 6 M urea, at neutral pH and 4°C. The purity of the proteins was analyzed by two‐dimensional gel electrophoresis. In addition, ten protein complexes were isolated and identified.