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Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands
Our fundamental understanding of proteins and their biological significance has been enhanced by genetic fusion tags, as they provide a convenient method for introducing unique properties to proteins so that they can be examinedin isolation. Commonly used tags satisfy many of the requirements for ap...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bentham Open
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3520037/ https://www.ncbi.nlm.nih.gov/pubmed/23248739 http://dx.doi.org/10.2174/1875397301206010055 |
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author | Encell, Lance P Friedman Ohana, Rachel Zimmerman, Kris Otto, Paul Vidugiris, Gediminas Wood, Monika G Los, Georgyi V McDougall, Mark G Zimprich, Chad Karassina, Natasha Learish, Randall D Hurst, Robin Hartnett, James Wheeler, Sarah Stecha, Pete English, Jami Zhao, Kate Mendez, Jacqui Benink, Hélène A Murphy, Nancy Daniels, Danette L Slater, Michael R Urh, Marjeta Darzins, Aldis Klaubert, Dieter H Bulleit, Robert F Wood, Keith V |
author_facet | Encell, Lance P Friedman Ohana, Rachel Zimmerman, Kris Otto, Paul Vidugiris, Gediminas Wood, Monika G Los, Georgyi V McDougall, Mark G Zimprich, Chad Karassina, Natasha Learish, Randall D Hurst, Robin Hartnett, James Wheeler, Sarah Stecha, Pete English, Jami Zhao, Kate Mendez, Jacqui Benink, Hélène A Murphy, Nancy Daniels, Danette L Slater, Michael R Urh, Marjeta Darzins, Aldis Klaubert, Dieter H Bulleit, Robert F Wood, Keith V |
author_sort | Encell, Lance P |
collection | PubMed |
description | Our fundamental understanding of proteins and their biological significance has been enhanced by genetic fusion tags, as they provide a convenient method for introducing unique properties to proteins so that they can be examinedin isolation. Commonly used tags satisfy many of the requirements for applications relating to the detection and isolation of proteins from complex samples. However, their utility at low concentration becomes compromised if the binding affinity for a detection or capture reagent is not adequate to produce a stable interaction. Here, we describe HaloTag® (HT7), a genetic fusion tag based on a modified haloalkane dehalogenase designed and engineered to overcome the limitation of affinity tags by forming a high affinity, covalent attachment to a binding ligand. HT7 and its ligand have additional desirable features. The tag is relatively small, monomeric, and structurally compatible with fusion partners, while the ligand is specific, chemically simple, and amenable to modular synthetic design. Taken together, the design features and molecular evolution of HT7 have resulted in a superior alternative to common tags for the overexpression, detection, and isolation of target proteins. |
format | Online Article Text |
id | pubmed-3520037 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Bentham Open |
record_format | MEDLINE/PubMed |
spelling | pubmed-35200372012-12-17 Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands Encell, Lance P Friedman Ohana, Rachel Zimmerman, Kris Otto, Paul Vidugiris, Gediminas Wood, Monika G Los, Georgyi V McDougall, Mark G Zimprich, Chad Karassina, Natasha Learish, Randall D Hurst, Robin Hartnett, James Wheeler, Sarah Stecha, Pete English, Jami Zhao, Kate Mendez, Jacqui Benink, Hélène A Murphy, Nancy Daniels, Danette L Slater, Michael R Urh, Marjeta Darzins, Aldis Klaubert, Dieter H Bulleit, Robert F Wood, Keith V Curr Chem Genomics Article Our fundamental understanding of proteins and their biological significance has been enhanced by genetic fusion tags, as they provide a convenient method for introducing unique properties to proteins so that they can be examinedin isolation. Commonly used tags satisfy many of the requirements for applications relating to the detection and isolation of proteins from complex samples. However, their utility at low concentration becomes compromised if the binding affinity for a detection or capture reagent is not adequate to produce a stable interaction. Here, we describe HaloTag® (HT7), a genetic fusion tag based on a modified haloalkane dehalogenase designed and engineered to overcome the limitation of affinity tags by forming a high affinity, covalent attachment to a binding ligand. HT7 and its ligand have additional desirable features. The tag is relatively small, monomeric, and structurally compatible with fusion partners, while the ligand is specific, chemically simple, and amenable to modular synthetic design. Taken together, the design features and molecular evolution of HT7 have resulted in a superior alternative to common tags for the overexpression, detection, and isolation of target proteins. Bentham Open 2012-10-05 /pmc/articles/PMC3520037/ /pubmed/23248739 http://dx.doi.org/10.2174/1875397301206010055 Text en ©Encell et al.; Licensee Bentham Open. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited. |
spellingShingle | Article Encell, Lance P Friedman Ohana, Rachel Zimmerman, Kris Otto, Paul Vidugiris, Gediminas Wood, Monika G Los, Georgyi V McDougall, Mark G Zimprich, Chad Karassina, Natasha Learish, Randall D Hurst, Robin Hartnett, James Wheeler, Sarah Stecha, Pete English, Jami Zhao, Kate Mendez, Jacqui Benink, Hélène A Murphy, Nancy Daniels, Danette L Slater, Michael R Urh, Marjeta Darzins, Aldis Klaubert, Dieter H Bulleit, Robert F Wood, Keith V Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title | Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title_full | Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title_fullStr | Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title_full_unstemmed | Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title_short | Development of a Dehalogenase-Based Protein Fusion Tag Capable of Rapid, Selective and Covalent Attachment to Customizable Ligands |
title_sort | development of a dehalogenase-based protein fusion tag capable of rapid, selective and covalent attachment to customizable ligands |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3520037/ https://www.ncbi.nlm.nih.gov/pubmed/23248739 http://dx.doi.org/10.2174/1875397301206010055 |
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