کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2048490 1074082 2012 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
N-Acetyl lysyl-tRNA synthetases evolved by a CcdB-based selection possess N-acetyl lysine specificity in vitro and in vivo
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک دانش گیاه شناسی
پیش نمایش صفحه اول مقاله
N-Acetyl lysyl-tRNA synthetases evolved by a CcdB-based selection possess N-acetyl lysine specificity in vitro and in vivo
چکیده انگلیسی

Posttranslational modifications play a crucial role in modulating protein structure and function. Genetic incorporation of unnatural amino acids into a specific site of a protein facilitates the systematic study of protein modifications including acetylation. We here report the directed evolution of pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei to create N-acetyl lysyl-tRNA synthetases (AcKRSs) using a new selection system based on the killing activity of the toxic ccdB gene product. The amino acid specificity of these and of published [1] and [2] AckRSs was tested in vitro and in vivo, and the enzyme-kinetic properties of the AckRSs were evaluated for the first time.


► N-acetyl lysyl-tRNA synthetases were evolved by a CcdB-based selection.
► N-acetyl lysine specificity was validated by both in vitro and in vivo approaches.
► Kinetic properties of the evolved synthetases were evaluated for the first time.
► The evolved synthetases will facilitate the systematic study of protein acetylation.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: FEBS Letters - Volume 586, Issue 6, 23 March 2012, Pages 729–733
نویسندگان
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