Crystal structures of leucyl/phenylalanyl-tRNA-protein transferase and its complex with an aminoacyl-tRNA analog

Kyoko Suto, Yoshihiro Shimizu, Kazunori Watanabe, Takuya Ueda, Shuya Fukai, Osamu Nureki, Kozo Tomita

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Eubacterial leucyl/phenylalanyl-tRNA protein transferase (L/F-transferase), encoded by the aat gene, conjugates leucine or phenylalanine to the N-terminal Arg or Lys residue of proteins, using Leu-tRNALeu or Phe-tRNA Phe as a substrate. The resulting N-terminal Leu or Phe acts as a degradation signal for the ClpS-ClpAP-mediated N-end rule protein degradation pathway. Here, we present the crystal structures of Escherichia coli L/F-transferase and its complex with an aminoacyl-tRNA analog, puromycin. The C-terminal domain of L/F-transferase consists of the GCN5-related N-acetyltransferase fold, commonly observed in the acetyltransferase superfamily. The p-methoxybenzyl group of puromycin, corresponding to the side chain of Leu or Phe of Leu-tRNALeu or Phe-tRNAPhe, is accommodated in a highly hydrophobic pocket, with a shape and size suitable for hydrophobic amino-acid residues lacking a branched β-carbon, such as leucine and phenylalanine. Structure-based mutagenesis of L/F-transferase revealed its substrate specificity. Furthermore, we present a model of the L/F-transferase complex with tRNA and substrate proteins bearing an N-terminal Arg or Lys.

Original languageEnglish
Pages (from-to)5942-5950
Number of pages9
JournalEMBO Journal
Volume25
Issue number24
DOIs
Publication statusPublished - 2006 Dec 13
Externally publishedYes

Fingerprint

leucyltransferase
Transferases
Transfer RNA
Crystal structure
RNA, Transfer, Leu
Puromycin
Acetyltransferases
Phenylalanine
Leucine
Bearings (structural)
Substrates
RNA, Transfer, Phe
Degradation
Mutagenesis
Proteins
Substrate Specificity
Escherichia coli
Proteolysis
Carbon
Genes

Keywords

  • Aminoacyl-tRNA
  • N-end rule
  • Protein stability
  • Transferase

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Crystal structures of leucyl/phenylalanyl-tRNA-protein transferase and its complex with an aminoacyl-tRNA analog. / Suto, Kyoko; Shimizu, Yoshihiro; Watanabe, Kazunori; Ueda, Takuya; Fukai, Shuya; Nureki, Osamu; Tomita, Kozo.

In: EMBO Journal, Vol. 25, No. 24, 13.12.2006, p. 5942-5950.

Research output: Contribution to journalArticle

Suto, Kyoko ; Shimizu, Yoshihiro ; Watanabe, Kazunori ; Ueda, Takuya ; Fukai, Shuya ; Nureki, Osamu ; Tomita, Kozo. / Crystal structures of leucyl/phenylalanyl-tRNA-protein transferase and its complex with an aminoacyl-tRNA analog. In: EMBO Journal. 2006 ; Vol. 25, No. 24. pp. 5942-5950.
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