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Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability
[Image: see text] For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793033/ https://www.ncbi.nlm.nih.gov/pubmed/29399652 http://dx.doi.org/10.1021/acsomega.7b01598 |
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author | Booth, William T. Schlachter, Caleb R. Pote, Swanandi Ussin, Nikita Mank, Nicholas J. Klapper, Vincent Offermann, Lesa R. Tang, Chuanbing Hurlburt, Barry K. Chruszcz, Maksymilian |
author_facet | Booth, William T. Schlachter, Caleb R. Pote, Swanandi Ussin, Nikita Mank, Nicholas J. Klapper, Vincent Offermann, Lesa R. Tang, Chuanbing Hurlburt, Barry K. Chruszcz, Maksymilian |
author_sort | Booth, William T. |
collection | PubMed |
description | [Image: see text] For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability. |
format | Online Article Text |
id | pubmed-5793033 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-57930332018-02-02 Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability Booth, William T. Schlachter, Caleb R. Pote, Swanandi Ussin, Nikita Mank, Nicholas J. Klapper, Vincent Offermann, Lesa R. Tang, Chuanbing Hurlburt, Barry K. Chruszcz, Maksymilian ACS Omega [Image: see text] For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability. American Chemical Society 2018-01-22 /pmc/articles/PMC5793033/ /pubmed/29399652 http://dx.doi.org/10.1021/acsomega.7b01598 Text en Copyright © 2018 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Booth, William T. Schlachter, Caleb R. Pote, Swanandi Ussin, Nikita Mank, Nicholas J. Klapper, Vincent Offermann, Lesa R. Tang, Chuanbing Hurlburt, Barry K. Chruszcz, Maksymilian Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability |
title | Impact of an N-terminal
Polyhistidine Tag on
Protein Thermal Stability |
title_full | Impact of an N-terminal
Polyhistidine Tag on
Protein Thermal Stability |
title_fullStr | Impact of an N-terminal
Polyhistidine Tag on
Protein Thermal Stability |
title_full_unstemmed | Impact of an N-terminal
Polyhistidine Tag on
Protein Thermal Stability |
title_short | Impact of an N-terminal
Polyhistidine Tag on
Protein Thermal Stability |
title_sort | impact of an n-terminal
polyhistidine tag on
protein thermal stability |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793033/ https://www.ncbi.nlm.nih.gov/pubmed/29399652 http://dx.doi.org/10.1021/acsomega.7b01598 |
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