PMID:17891155
| Citation |
Watanabe, K, Toh, Y, Suto, K, Shimizu, Y, Oka, N, Wada, T and Tomita, K (2007) Protein-based peptide-bond formation by aminoacyl-tRNA protein transferase. Nature 449:867-71 |
|---|---|
| Abstract |
Eubacterial leucyl/phenylalanyl-tRNA protein transferase (LF-transferase) catalyses peptide-bond formation by using Leu-tRNA(Leu) (or Phe-tRNA(Phe)) and an amino-terminal Arg (or Lys) of a protein, as donor and acceptor substrates, respectively. However, the catalytic mechanism of peptide-bond formation by LF-transferase remained obscure. Here we determine the structures of complexes of LF-transferase and phenylalanyl adenosine, with and without a short peptide bearing an N-terminal Arg. Combining the two separate structures into one structure as well as mutation studies reveal the mechanism for peptide-bond formation by LF-transferase. The electron relay from Asp 186 to Gln 188 helps Gln 188 to attract a proton from the alpha-amino group of the N-terminal Arg of the acceptor peptide. This generates the attacking nucleophile for the carbonyl carbon of the aminoacyl bond of the aminoacyl-tRNA, thus facilitating peptide-bond formation. The protein-based mechanism for peptide-bond formation by LF-transferase is similar to the reverse reaction of the acylation step observed in the peptide hydrolysis reaction by serine proteases. |
| Links |
PubMed Online version:10.1038/nature06167 |
| Keywords |
Acylation; Adenosine/analogs & derivatives; Adenosine/metabolism; Amino Acid Sequence; Binding Sites; Caseins/biosynthesis; Caseins/chemistry; Catalysis; Escherichia coli/enzymology; Hydrophobic and Hydrophilic Interactions; Kinetics; Models, Molecular; Peptide Fragments/biosynthesis; Peptide Fragments/chemistry; Peptidyl Transferases/chemistry; Peptidyl Transferases/metabolism; Phenylalanine/metabolism; Phenylalanine-tRNA Ligase/metabolism; Protein Biosynthesis; Protein Conformation; Puromycin/chemistry; Puromycin/metabolism; RNA, Transfer, Amino Acyl/chemistry; RNA, Transfer, Amino Acyl/metabolism; Substrate Specificity |
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