415215 Thiazoline Ring Formation in an Engineered Intein Serves As a Splicing Switch

Tuesday, November 10, 2015: 9:45 AM
150G (Salt Palace Convention Center)
C. Seth Pearson1, Dan Fabris2, Hongmin Li3, Georges Belfort4 and Marlene Belfort2, (1)Howard P. Isermann Dept of Chemical & Biological Engineering and The Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, (2)University at Albany, Albany, NY, (3)Wadsworth Center, Albany, NY, (4)Howard P. Isermann Dept of Chemical & Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY

We have engineered an intein which autocatalytically forms a thiazoline ring at the N-terminal splicing junction and provided the first crystallographic evidence of this chemistry in an intein. The mutation that causes the aberrant thiazoline ring chemistry suggests that the highly conserved B-block Thr plays a pivotal role in orienting the resulting tetrahedral intermediate for resolution into a thioester, giving insight into the splicing mechanism. We demonstrate the stability of the thiazoline ring at mild conditions as well as sensitivity to hydrolytic opening of the ring under acidic conditions and high temperature. The reversibility of this thiazoline “trap” is of significant interest to biotechnological applications requiring a splicing activity switch.

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See more of this Session: Biomolecular Engineering
See more of this Group/Topical: Food, Pharmaceutical & Bioengineering Division