Expression and characterization of bovine mitochondrial methionyl-tRNA transformylase

Nono Takeuchi, Takuya Ueda, Kimitsuna Watanabe

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Translational initiation in bacteria and some organelles such as mitochondria and chloroplasts requires formyl-methionyl-tRNA (fMet-tRNA). Methionyl-tRNA (Met-tRNA) undergoes formylation by methionyl-tRNA transformylase (MTF), and the resulting fMet-tRNA is utilized exclusively in the initiation process. The gene encoding mammalian mitochondrial MTF (MTFmt) was cloned recently. When the cDNA corresponding to mature MTFmt was cloned into an expression vector, no expression of MTFmt was observed. However, if the cDNA was fused with the histidine-tag sequence at the N-terminus, MTFmt could be expressed in Escherichia coli. The recombinant enzyme was purified by a single step on a histidine-binding metal affinity column. We previously found that native MTFmt is able to formylate E. coli elongator Met-tRNA as well as the initiator Met-tRNA. The specific formylation of the initiator Met-tRNA by E. coli MTF is quite important in bacterial translational initiation. The purified recombinant MTFmt with the histidine-tag showed almost identical kinetic parameters to those of native MTFmt. This expression system is suitable for the rapid, efficient production of MTFmt in amounts adequate for further biophysical studies, which will provide another approach for elucidating the formylation mechanism, in addition to studies on E. coli MTF.

Original languageEnglish
Pages (from-to)1069-1071
Number of pages3
JournalJournal of biochemistry
Volume124
Issue number6
DOIs
Publication statusPublished - 1998 Jan 1
Externally publishedYes

Fingerprint

methionyl-tRNA formyltransferase
Transfer RNA
Escherichia coli
Histidine
RNA, Transfer, Met
Complementary DNA
Chloroplasts
Mitochondria
Gene encoding
Organelles
Kinetic parameters
Metals
Bacteria
Enzymes
Genes

Keywords

  • Expression
  • Mammalian mitochondria
  • MTF
  • Substrate specificity
  • Translational initiation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Expression and characterization of bovine mitochondrial methionyl-tRNA transformylase. / Takeuchi, Nono; Ueda, Takuya; Watanabe, Kimitsuna.

In: Journal of biochemistry, Vol. 124, No. 6, 01.01.1998, p. 1069-1071.

Research output: Contribution to journalArticle

Takeuchi, Nono ; Ueda, Takuya ; Watanabe, Kimitsuna. / Expression and characterization of bovine mitochondrial methionyl-tRNA transformylase. In: Journal of biochemistry. 1998 ; Vol. 124, No. 6. pp. 1069-1071.
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