원문정보
초록
영어
Protein trans-splicing reaction (PTS) is self-processing enzymatic reaction mediated by a pair of split-inteins. PTS based protein semi-synthesis enables the introduction of various synthetic probes to target proteins in vivo and in vitro. While the PTS became a useful tool for various biological studies, the difficulties associated with the synthesis of 40 mer peptide became a drawback in its applications. Here we have generated several engineered split-inteins by shifting the split-site of naturally split-inteins to new positions. We utilized Npu DnaE intein as a model system as this naturally split-intein mediates fast (t1/2 ~ 1 min) PTS in physiological conditions. In order to investigate how the lengths of N and C intein affect the PTS kinetics, we prepared multiple pairs of split-inteins that consist of various lengths of N and C-inteins. The kinetics of reaction between each engineered pairs and the cross-reactivities were studied.