Construction of modified ribosomes for incorporation of D-amino acids into proteins

Larisa M. Dedkova, Nour Eddine Fahmi, Serguei Y. Golovine, Sidney M. Hecht

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

While numerous biologically active peptides contain D-amino acids, the elaboration of such species is not carried out by ribosomal synthesis. In fact, the bacterial ribosome discriminates strongly against the incorporation of D-amino acids from D-aminoacyl-tRNAs. To permit the incorporation of D-amino acids into proteins using in vitro protein-synthesizing systems, a strategy has been developed to prepare modified ribosomes containing alterations within the peptidyltransferase center and helix 89 of 23S rRNA. S-30 preparations derived from colonies shown to contain ribosomes with altered 23S rRNAs were found to exhibit enhanced tolerance for D-amino acids and to permit the elaboration of proteins containing D-amino acids at predetermined sites. Five specific amino acids in Escherichia coli dihydrofolate reductase and Photinus pyralis luciferase were replaced with D-phenylalanine and D-methionine, and the specific activities of the resulting enzymes were determined.

Original languageEnglish (US)
Pages (from-to)15541-15551
Number of pages11
JournalBiochemistry
Volume45
Issue number51
DOIs
StatePublished - Dec 26 2006
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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