| RRC ID |
35638
|
| Author |
Fung AW, Ebhardt HA, Krishnakumar KS, Moore J, Xu Z, Strazewski P, Fahlman RP.
|
| Title |
Probing the leucyl/phenylalanyl tRNA protein transferase active site with tRNA substrate analogues.
|
| Journal |
Protein Pept Lett
|
| Abstract |
Aminoacyl-tRNA protein transferases post-translationally conjugate an amino acid from an aminoacyl-tRNA onto the N-terminus of a target polypeptide. The eubacterial aminoacyl-tRNA protein transferase, L/F transferase, utilizes both leucyl-tRNA(Leu) and phenylalanyl-tRNA(Phe) as substrates. X-ray crystal structures with substrate analogues, the minimal substrate phenylalanyl adenosine (rA-Phe) and inhibitor puromycin, have been used to characterize tRNA recognition by L/F transferase. However analyses of these two X-ray crystal structures reveal significant differences in binding. Through structural analyses, mutagenesis, and enzymatic activity assays, we rationalize and demonstrate that the substrate analogues bind to L/F transferase with similar binding affinities using a series of different interactions by the various chemical groups of the analogues. Our data also demonstrates that enlarging the hydrophobic pocket of L/F transferase selectively enhances puromycin inhibition and may aid in the development of improved inhibitors for this class of enzymes.
|
| Volume |
21(7)
|
| Pages |
603-14
|
| Published |
2014-7-1
|
| DOI |
10.2174/0929866521666140212110639
|
| PII |
PPL-EPUB-59140
|
| PMID |
24521222
|
| MeSH |
Aminoacyltransferases* / antagonists & inhibitors
Aminoacyltransferases* / chemistry
Aminoacyltransferases* / metabolism
Catalytic Domain
Crystallography, X-Ray
Escherichia coli Proteins
Mass Spectrometry
Models, Molecular
Mutation
Protein Binding
Puromycin / pharmacology
RNA, Transfer, Leu / chemistry*
RNA, Transfer, Leu / metabolism*
RNA, Transfer, Phe / chemistry*
RNA, Transfer, Phe / metabolism*
Recombinant Fusion Proteins
|
| IF |
1.156
|
| Times Cited |
3
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Prokaryotes E. coli |
NA |
