Identification of the N-tosyl-L-phenylalanyl chloromethylketone modification site in Thermus thermophilus elongation factor Tu

Marcus E. Peter*, Jürgen Brockmöller, Jiří Jonák, Mathias Sprinzl

*Corresponding author for this work

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

EF-Tu from Thermus thermophilus was first labelled with N-[14C]tosyl-L-phenylalaninechloromethylketone and then cleaved by the combined action of CNBr and trypsin. The resulting peptides were separated by reversed-phase HPLC. Analysis of the isolated, labelled peptide led to the identification of a sequence which was identical to residues 76-88 in T. thermophilus EF-Tu. The TPCK reactive site is at Cys-82. Kinetic measurements f the incorporation of TPCK into native EF-Tu and EF-Tu nicked at position Arg-59 were performed. The results provide evidence that the cleavage of the peptide bond between Arg-59 and Gly-60 does not lead to a dramatic conformational change of EF-Tu at the aa-tRNA binding site.

Original languageEnglish (US)
Pages (from-to)219-222
Number of pages4
JournalFEBS Letters
Volume257
Issue number2
DOIs
StatePublished - Nov 6 1989

Keywords

  • (Thermus thermophilus)
  • Elongation factor Tu
  • GTP-binding protein
  • N-Tosyl-L-phenylalanyl chloromethylketone
  • Protein biosynthesis

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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