Ehrlich Michael, Gattner Michael J, Viverge Bastien, Bretzler Johanna, Eisen David, Stadlmeier Michael, Vrabel Milan, Carell Thomas
Center for Integrative Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, Butenandtstrasse 5-13, 81377 Munich (Germany).
Chemistry. 2015 May 18;21(21):7701-4. doi: 10.1002/chem.201500971. Epub 2015 Apr 2.
We here report the construction of an E. coli expression system able to manufacture an unnatural amino acid by artificial biosynthesis. This can be orchestrated with incorporation into protein by amber stop codon suppression inside a living cell. In our case an alkyne-bearing pyrrolysine amino acid was biosynthesized and incorporated site-specifically allowing orthogonal double protein labeling.
我们在此报告构建了一种大肠杆菌表达系统,该系统能够通过人工生物合成制造一种非天然氨基酸。这可以通过在活细胞内利用琥珀色终止密码子抑制将其掺入蛋白质中来精心安排。在我们的例子中,一种带有炔基的吡咯赖氨酸氨基酸被生物合成并位点特异性地掺入,从而实现正交双蛋白标记。