| Autor: |
Wagner AM; Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6323, United States., Fegley MW, Warner JB, Grindley CL, Marotta NP, Petersson EJ |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of the American Chemical Society [J Am Chem Soc] 2011 Sep 28; Vol. 133 (38), pp. 15139-47. Date of Electronic Publication: 2011 Sep 06. |
| DOI: |
10.1021/ja2055098 |
| Abstrakt: |
Methods for synthetically manipulating protein structure enable greater flexibility in the study of protein function. Previous characterization of the Escherichia coli aminoacyl tRNA transferase (AaT) has shown that it can modify the N-terminus of a protein with an amino acid from a tRNA or a synthetic oligonucleotide donor. Here, we demonstrate that AaT can efficiently use a minimal adenosine substrate, which can be synthesized in one to two steps from readily available starting materials. We have characterized the enzymatic activity of AaT with aminoacyl adenosyl donors and found that reaction products do not inhibit AaT. The use of adenosyl donors removes the substrate limitations imposed by the use of synthetases for tRNA charging and avoids the complex synthesis of an oligonucleotide donor. Thus, our AaT donors increase the potential substrate scope and reaction scale for N-terminal protein modification under conditions that maintain folding. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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