Structural studies on tubulin tyrosine ligase and its application.

我们建立了E。大肠杆菌表达系统中表达全长人微管蛋白酪氨酸连接酶1（TTL1）。利用共表达技术，共表达了分子伴侣，提高了TTL1的溶解性，成功地将产量提高了约5倍。我们还找到了核磁共振测量的最佳缓冲条件。我们还开发了使用GFP融合的TTL1的高通量监测系统。同时，我们建立了TTL1活性的核磁共振实时监测系统。

We have established the E. coli expression system for the full-length human tubulin tyrosine ligase 1 (TTL1). By using co-expression technique, molecular chaperone was co-expressed to improve the solubility of TTL1, and we succeeded in improvement of the yield about 5 times. We also found that the optimal buffer condition for NMR measurement. We have also developed the high-throughput monitoring system using GFP fused TTL1. Meanwhile, we have established the real-time monitoring system of TTL1 activity by NMR.