Nε-acetyllysine transfer ribonucleic acid

A biologically active analogue of aminoacyl transfer ribonucleic acids

Arthur E. Johnson, William Woodward, Edward Herbert, John R. Menninger

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Unfractionated Escherichia coli tRNA has been aminoacylated with lysine and preferentially acetylated at the ε-amino nitrogen of lysine by reaction with N-acetoxysuccinimide. After treatment with peptidyl-tRNA hydrolase, 90% of the aminoacylated tRNA molecules were Nε-acetyl-Lys-tRNA. Post-ribosomal supernatant enzymes would not deacylate Nε-acetyl-Lys-tRNA in the presence of AMP and PPi, even though such mixed enzymes could acylate, with lysine, tRNA which had been exposed to the acetylation reaction conditions. Poly(rA) stimulated the binding of Nε-acetyl-Lys-tRNA to E. coli ribosomes. At the ribosome and tRNA concentrations used, Nε-acetyl-Lys-tRNA was bound nearly as well as Lys-tRNA at 30 mM Mg2+; at 10 mM Mg2+, the analogue was bound one-half as well as Lys-tRNA. Both Lys-tRNA and Nε-acetyl-Lys-tRNA reacted only slightly with puromycin at either 10 or 30 mM Mg2+. When Lys-tRNAE. coli or Nε-acetyl-Lys-tRNAE. coli were added to rabbit reticulocyte cell-free protein synthesizing incubations, the incorporation of either amino acid into protein was complete within 5 min. The final incorporation level of the analogue was 82% that of the unmodified lysine. After protein synthesized in the presence of Nε-acetyl-[14C]Lys-tRNA had been digested enzymatically to single amino acids, ion-exchange chromatography and paper electrophoresis showed that nearly all of the radioactivity was present as Nε-acetyllysine. Gel filtration of the post-ribosomal supernatant revealed that most of the Nε-acetyllysine radioactivity cochromatographed with tetrameric hemoglobin.

Original languageEnglish (US)
Pages (from-to)569-575
Number of pages7
JournalBiochemistry
Volume15
Issue number3
StatePublished - 1976
Externally publishedYes

Fingerprint

RNA, Transfer, Lys
Transfer RNA
Lysine
Radioactivity
Ribosomes
Escherichia coli
Paper Electrophoresis
Amino Acids
Acetylation
Puromycin
Proteins
Poly A
Reticulocytes
Ion Exchange Chromatography
Enzymes
Adenosine Monophosphate
Chromatography
Electrophoresis
Gel Chromatography
Ion exchange

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nε-acetyllysine transfer ribonucleic acid : A biologically active analogue of aminoacyl transfer ribonucleic acids. / Johnson, Arthur E.; Woodward, William; Herbert, Edward; Menninger, John R.

In: Biochemistry, Vol. 15, No. 3, 1976, p. 569-575.

Research output: Contribution to journalArticle

Johnson, Arthur E. ; Woodward, William ; Herbert, Edward ; Menninger, John R. / Nε-acetyllysine transfer ribonucleic acid : A biologically active analogue of aminoacyl transfer ribonucleic acids. In: Biochemistry. 1976 ; Vol. 15, No. 3. pp. 569-575.
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