RRC ID 35638
著者 Fung AW, Ebhardt HA, Krishnakumar KS, Moore J, Xu Z, Strazewski P, Fahlman RP.
タイトル Probing the leucyl/phenylalanyl tRNA protein transferase active site with tRNA substrate analogues.
ジャーナル 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.
巻・号 21(7)
ページ 603-14
公開日 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
引用数 3
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
原核生物(大腸菌) NA