Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul, South Korea.
FEBS Lett. 2011 Jul 21;585(14):2269-74. doi: 10.1016/j.febslet.2011.05.051. Epub 2011 May 27.
We found that the synthesis of histone H3 N-terminal peptide (tail) in a reconstituted protein synthesis system yielded fragmented peptides along with the full-length product. With the combined use of MALDI-TOF analysis and peptidyl-tRNA hydrolase cleavage of the Flag tagged product species, we concluded that the fragments were generated by peptidyl-tRNA drop-off at specific sites and subsequent translation continuation. Using the histone H3 tail we also found that peptidyl-tRNA drop-off is strongly correlated with the amino acid context. We envision that the system described here would be useful as a model system for studying peptidyl-tRNA drop-off events.
我们发现，在重新构建的蛋白质合成体系中合成组蛋白 H3 N 端肽（尾部）会产生全长产物的同时生成一些片段肽。通过 MALDI-TOF 分析和 Flag 标记产物的肽基-tRNA 水解酶切割的联合使用，我们得出结论，这些片段是由特定位置的肽基-tRNA 脱落和随后的翻译延伸产生的。使用组蛋白 H3 尾部，我们还发现肽基-tRNA 脱落与氨基酸序列上下文密切相关。我们设想，这里描述的体系将作为研究肽基-tRNA 脱落事件的模型体系非常有用。
