Ribosomal synthesis of polylysine from individual lysyl-tRNA/sup Lys/ in the absence of a template

Journal Article · Thu Jan 01 04:00:00 EST 1987 · Dokl. Biochem. (Engl. Transl.); (United States)

OSTI ID:5856593

Yusupova, G Z; Remme, Y L; Belitsina, N B; Spirin, A S

Earlier studies showed that ribosomes of Escherichia coli, in the absence of a template, can synthesize oligolysine, using lysyl-tRNA as a substrate. The authors present results on the use of preparations of individual lysyl-tRNA/sup Lys/ and phenylalanyl-tRNA/sup Phe/ in a system of templateless peptide synthesis. For these studies, the authors used ribosomes of E. coli MRE 600, washed four times with 1 M NH/sub 4/Cl with 10 MM MgCl/sub 2/. The purified ribosomes were stored at -70/sup 0/C in standard buffer, containing 20 mM Tris-HCl, 100 mM NH/sub 4/Cl, 10 mM MgCl/sub 2/, 0.1 mM ethylenediamine tetraacetate (EDTA), and 10% glycerin, pH/sub 37/sup 0/C/7.6. A preparation of (/sup 14/C)lysyl-tRNA/sup Lys/ was produced by affinity chromatography on immobilized factor EF-T/sub u/ from Thermus thermophilus HB8. The elongation factor EF-T/sub u/ from T. thermophilus and immobilized on BrCN-activated Sepharose 4B. The initial preparation of total tRNA of E. coli, enzymatically acylated by (/sup 14/C)lysine (348 Ci/mole, Amersham), was produced as described earlier. The degree of aminoacylation was 52-59 pmoles (/sup 14/C)lysine per unit of A/sub 260/ of tRNA.

• Research Organization: Protein Institute, Pushchino, USSR
• OSTI ID: 5856593
• Journal Information: Dokl. Biochem. (Engl. Transl.); (United States), Journal Name: Dokl. Biochem. (Engl. Transl.); (United States) Vol. 289:1-6; ISSN DBIOA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACTIVATION ENERGY
ACYLATION
AMINATION
AMINO ACIDS
BACTERIA
BIOCHEMICAL REACTION KINETICS
BIOCHEMISTRY
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHEMISTRY
ELONGATION
ENERGY
ESCHERICHIA COLI
KINETICS
LABELLED COMPOUNDS
LABELLING
LYSINE
METABOLIC ACTIVATION
METABOLISM
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANOIDS
POLYMERIZATION
RADIOENZYMATIC ASSAY
REACTION KINETICS
RIBOSOMES
RNA
SYNTHESIS