Abstract
RNase HI from the hyperthermophile Sulfolobus tokodaii (Sto-RNase HI) is stabilized by its C-terminal residues. In this work, the stabilization effect of the Sto-RNase HI C-terminal residues was investigated in detail by thermodynamic measurements of the stability of variants lacking the disulfide bond (C58/145A), or the six C-terminal residues (ΔC6) and by structural analysis of ΔC6. The results showed that the C-terminal does not affect overall structure and stabilization is caused by local interactions of the C-terminal, suggesting that the C-terminal residues could be used as a “stabilization tag.” The Sto-RNase HI C-terminal residues (-IGCIILT) were introduced as a tag on three proteins. Each chimeric protein was more stable than its wild-type protein. These results suggested the possibility of a simple stabilization technique using a stabilization tag such as Sto-RNase HI C-terminal residues.
Highlights
An important goal of protein engineering is designing variants that enhance the conformational stability of proteins
Protein tags, which are peptide sequences genetically grafted onto the N- or C-terminus of recombinant proteins, are widely used experimentally because they are easy to manipulate
The C-terminus of Sto-RNase HI is anchored to the core region by one disulfide bond (Cys58-Cys145), several hydrogen bonds, and hydrophobic interactions (Figure 1)
Summary
An important goal of protein engineering is designing variants that enhance the conformational stability of proteins. We analyzed stabilization by the amino acid residues of the Sto-RNase HI C-terminus, and aimed to develop a protein stabilization tag. We investigated in detail the stabilization effect of the Sto-RNase HI C-terminus, by measuring the stability of a C58/145A variant lacking the disulfide bond, and a DC6 variant lacking the six C-terminal residues. The Sto-RNase HI C-terminal residues IGCIILT were introduced onto three proteins as a tag, and the effect on stability was examined.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
