T4 RNA Ligase Mediated Preparation of Novel “Chemically Misacylated” tRNAphes

Thomas G. Heckler, Li Ho Chang, Yoshiyuki Zama, Takehiko Naka, Mukund S. Chorghade, Sidney M. Hecht

Research output: Contribution to journalArticlepeer-review

163 Scopus citations

Abstract

T4 RNA ligase was employed for the condensation of Escherichia coli tRNAPhe missing cytidine-75 and adenosine-76 (tRNAPhe-COH; the acceptor “oligomer”) with each of several chemically acylated derivatives of pCpA”(the donor oligomer). The resulting chemically misacylated tRNA s were obtained in 20-65% yields following chromatographic workup on DEAE-cellulose and benzoylated DEAE-cellulose. Characterization of the chemically misacylated tRNAs was accomplished by (i) enzymatic reaminoacylation of chemically misacylated tRNA with phenylalanine by E. coli phenylalanyl-tRNA synthetase following chemical deacylation of the “incorrect” amino acid, (ii) comparison of the hydrolytic effects of Cu2+ solutions on chemically and enzymatically prepared samples of N-acetyl-L-phenylalanyl-tRNAPhes, and (iii) measurement of the chromatographic behavior of the tRNA species derived from chemical misacylation.

Original languageEnglish (US)
Pages (from-to)1468-1473
Number of pages6
JournalBiochemistry
Volume23
Issue number7
DOIs
StatePublished - Mar 1984
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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