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Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag

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 b...

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Autores principales: Takano, Kazufumi, Okamoto, Tomohiro, Okada, Jun, Tanaka, Shun-ichi, Angkawidjaja, Clement, Koga, Yuichi, Kanaya, Shigenori
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3023800/
https://www.ncbi.nlm.nih.gov/pubmed/21283826
http://dx.doi.org/10.1371/journal.pone.0016226
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author Takano, Kazufumi
Okamoto, Tomohiro
Okada, Jun
Tanaka, Shun-ichi
Angkawidjaja, Clement
Koga, Yuichi
Kanaya, Shigenori
author_facet Takano, Kazufumi
Okamoto, Tomohiro
Okada, Jun
Tanaka, Shun-ichi
Angkawidjaja, Clement
Koga, Yuichi
Kanaya, Shigenori
author_sort Takano, Kazufumi
collection PubMed
description 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.
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spelling pubmed-30238002011-01-31 Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag Takano, Kazufumi Okamoto, Tomohiro Okada, Jun Tanaka, Shun-ichi Angkawidjaja, Clement Koga, Yuichi Kanaya, Shigenori PLoS One Research Article 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. Public Library of Science 2011-01-19 /pmc/articles/PMC3023800/ /pubmed/21283826 http://dx.doi.org/10.1371/journal.pone.0016226 Text en Takano et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Takano, Kazufumi
Okamoto, Tomohiro
Okada, Jun
Tanaka, Shun-ichi
Angkawidjaja, Clement
Koga, Yuichi
Kanaya, Shigenori
Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title_full Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title_fullStr Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title_full_unstemmed Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title_short Stabilization by Fusion to the C-terminus of Hyperthermophile Sulfolobus tokodaii RNase HI: A Possibility of Protein Stabilization Tag
title_sort stabilization by fusion to the c-terminus of hyperthermophile sulfolobus tokodaii rnase hi: a possibility of protein stabilization tag
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3023800/
https://www.ncbi.nlm.nih.gov/pubmed/21283826
http://dx.doi.org/10.1371/journal.pone.0016226
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