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Rapid purification of HU protein from Halobacillus karajensis

The histone-like protein HU is the most-abundant DNA-binding protein in bacteria. The HU protein non-specifically binds and bends DNA as a hetero- or homodimer, and can participate in DNA supercoiling and DNA condensation. It also takes part in DNA functions such as replication, recombination, and r...

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Detalles Bibliográficos
Autores principales: Ghadam, Parinaz, Samadi, Rana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Shiraz University 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5019216/
https://www.ncbi.nlm.nih.gov/pubmed/27843972
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author Ghadam, Parinaz
Samadi, Rana
author_facet Ghadam, Parinaz
Samadi, Rana
author_sort Ghadam, Parinaz
collection PubMed
description The histone-like protein HU is the most-abundant DNA-binding protein in bacteria. The HU protein non-specifically binds and bends DNA as a hetero- or homodimer, and can participate in DNA supercoiling and DNA condensation. It also takes part in DNA functions such as replication, recombination, and repair. HU does not recognize any specific sequences but shows a certain degree of specificity to cruciform DNA and repair intermediates such as nick, gap, bulge, etc. To understand the features of HU binding to DNA and repair intermediates, a fast and easy HU protein purification method is required. Here we report a two-step purification procedure of HU from Halobacillus karajensis (the gram positive and moderately halophilic bacteria isolated from Karaj surface soil). The method of HU purification allows obtaining a pure non-tagged protein. Salting out and ion exchange chromatography were applied for purification, and the purified protein was identified by immunoblotting. Results showed that the molecular weight of the purified protein was approximately 11 kDa which is immunologically similar to the Bacillus subtilis HU protein (HBsu).
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spelling pubmed-50192162016-11-14 Rapid purification of HU protein from Halobacillus karajensis Ghadam, Parinaz Samadi, Rana Mol Biol Res Commun Original Article The histone-like protein HU is the most-abundant DNA-binding protein in bacteria. The HU protein non-specifically binds and bends DNA as a hetero- or homodimer, and can participate in DNA supercoiling and DNA condensation. It also takes part in DNA functions such as replication, recombination, and repair. HU does not recognize any specific sequences but shows a certain degree of specificity to cruciform DNA and repair intermediates such as nick, gap, bulge, etc. To understand the features of HU binding to DNA and repair intermediates, a fast and easy HU protein purification method is required. Here we report a two-step purification procedure of HU from Halobacillus karajensis (the gram positive and moderately halophilic bacteria isolated from Karaj surface soil). The method of HU purification allows obtaining a pure non-tagged protein. Salting out and ion exchange chromatography were applied for purification, and the purified protein was identified by immunoblotting. Results showed that the molecular weight of the purified protein was approximately 11 kDa which is immunologically similar to the Bacillus subtilis HU protein (HBsu). Shiraz University 2014-03 /pmc/articles/PMC5019216/ /pubmed/27843972 Text en This is an Open Access article distributed under the terms of the Creative Commons Attribution License, (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Ghadam, Parinaz
Samadi, Rana
Rapid purification of HU protein from Halobacillus karajensis
title Rapid purification of HU protein from Halobacillus karajensis
title_full Rapid purification of HU protein from Halobacillus karajensis
title_fullStr Rapid purification of HU protein from Halobacillus karajensis
title_full_unstemmed Rapid purification of HU protein from Halobacillus karajensis
title_short Rapid purification of HU protein from Halobacillus karajensis
title_sort rapid purification of hu protein from halobacillus karajensis
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5019216/
https://www.ncbi.nlm.nih.gov/pubmed/27843972
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