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. 1968 Apr;95(4):1355–1367. doi: 10.1128/jb.95.4.1355-1367.1968

Identification of a Precursor Pool of Ribosome Protein in Escherichia coli

Melvin Santer a, Trenton K Ruebush a,1, John Van Brunt a, Eben Oldmixon a,2, Russell Hess a,3, Paul Primakoff a,4, Philip Palade a,5
PMCID: PMC315094  PMID: 4967196

Abstract

Antibodies prepared against proteins from 50S ribosomes of Escherichia coli also reacted with the supernatant proteins of a cell-free extract of E. coli which was ribosome-free. A reaction of immunological identity (Ouchterlony tests) was demonstrated for one of these supernatant proteins and one protein found in 50S ribosomes. Isotope experiments involving a shift from 14C-leucine medium to 12C-leucine medium showed that these proteins are not formed by breakdown of ribosomes during the preparation of cell-free extracts, but instead represent a pool of ribosome protein which is utilized during growth. In shift experiments from 14C-leucine to 12C-leucine medium, the kinetics of disappearance of labeled supernatant ribosome proteins (as measured by reaction with antibody) indicated that half the pool is depleted in 0.1 generation time at 37 C in glucose-salts medium. The pool was also depleted under conditions of amino acid starvation of a “relaxed” strain which accumulated “relaxed” particles. Most, if not all, of the protein present in “relaxed” particles was derived from the pool. The pool represented about 3 to 4% of the total soluble proteins in the ribosome-free supernatant fluid of an E. coli extract.

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