Structural Analysis of Halophilic and Thermophilic Inteins

Abstract

Protein splicing is a post‐translational modification by which a segment of protein, known as an intein, self‐catalyzes its excision from the flanking polypeptides while simultaneously ligating the exteins. Inteins can be found in many types of archaebacteria, including halophiles, which thrive in environments with high salt concentrations. Two halophilic inteins that interrupt the Cdc21 gene, CdcA and CdcD, have been shown to splice at different salt concentrations. In addition, the PolII intein from the extreme thermophile Pyrococcus horikoshii requires incubation at high temperature in order to splice. We are interested in solving the structure of all three inteins by NMR to better probe the relationship between a protein's structure and its requirements of extreme conditions for activity. To obtain the proteins at sufficiently high concentrations for structural analysis, we sub‐cloned the inteins of interest into expression vectors with an N‐terminal GB1 tag and optimized expression conditions.Support or Funding InformationThis work was supported by the National Science Foundation (MCB‐1517138, KVM) and the Dreyfus Foundation (KVM).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.