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An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease

A putative protease gene (tldD) was previously identified from studying tolerance of letD encoding the CcdB toxin of a toxin–antidote system of the F plasmid in Escherichia coli. While this gene is evolutionarily conserved in archaea and bacteria, the proteolytic activity of encoded proteins remaine...

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Autores principales: Hu, Yongmei, Peng, Nan, Han, Wenyuan, Mei, Yuxia, Chen, Zhengjun, Feng, Xu, Liang, Yun Xiang, She, Qunxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497727/
https://www.ncbi.nlm.nih.gov/pubmed/22950735
http://dx.doi.org/10.1042/BSR20120074
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author Hu, Yongmei
Peng, Nan
Han, Wenyuan
Mei, Yuxia
Chen, Zhengjun
Feng, Xu
Liang, Yun Xiang
She, Qunxin
author_facet Hu, Yongmei
Peng, Nan
Han, Wenyuan
Mei, Yuxia
Chen, Zhengjun
Feng, Xu
Liang, Yun Xiang
She, Qunxin
author_sort Hu, Yongmei
collection PubMed
description A putative protease gene (tldD) was previously identified from studying tolerance of letD encoding the CcdB toxin of a toxin–antidote system of the F plasmid in Escherichia coli. While this gene is evolutionarily conserved in archaea and bacteria, the proteolytic activity of encoded proteins remained to be demonstrated experimentally. Here we studied Sso0660, an archaeal TldD homologue encoded in Sulfolobus solfataricus by overexpression of the recombinant protein and characterization of the purified enzyme. We found that the enzyme is active in degrading azocasein and FITC–BSA substrates. Protease inhibitor studies showed that EDTA and o-phenanthroline, two well-known metalloprotease inhibitors, either abolished completely or strongly inhibited the enzyme activity, and flame spectrometric analysis showed that a zinc ion is a cofactor of the protease. Furthermore, the protein forms disulfide bond via the Cys(416) residue, yielding protein dimer that is the active form of the enzyme. These results establish for the first time that tidD genes encode zinc-containing proteases, classifying them as a family in the metalloprotease class.
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spelling pubmed-34977272012-12-01 An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease Hu, Yongmei Peng, Nan Han, Wenyuan Mei, Yuxia Chen, Zhengjun Feng, Xu Liang, Yun Xiang She, Qunxin Biosci Rep Original Paper A putative protease gene (tldD) was previously identified from studying tolerance of letD encoding the CcdB toxin of a toxin–antidote system of the F plasmid in Escherichia coli. While this gene is evolutionarily conserved in archaea and bacteria, the proteolytic activity of encoded proteins remained to be demonstrated experimentally. Here we studied Sso0660, an archaeal TldD homologue encoded in Sulfolobus solfataricus by overexpression of the recombinant protein and characterization of the purified enzyme. We found that the enzyme is active in degrading azocasein and FITC–BSA substrates. Protease inhibitor studies showed that EDTA and o-phenanthroline, two well-known metalloprotease inhibitors, either abolished completely or strongly inhibited the enzyme activity, and flame spectrometric analysis showed that a zinc ion is a cofactor of the protease. Furthermore, the protein forms disulfide bond via the Cys(416) residue, yielding protein dimer that is the active form of the enzyme. These results establish for the first time that tidD genes encode zinc-containing proteases, classifying them as a family in the metalloprotease class. Portland Press Ltd. 2012-10-15 2012-12-01 /pmc/articles/PMC3497727/ /pubmed/22950735 http://dx.doi.org/10.1042/BSR20120074 Text en © 2012 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited
spellingShingle Original Paper
Hu, Yongmei
Peng, Nan
Han, Wenyuan
Mei, Yuxia
Chen, Zhengjun
Feng, Xu
Liang, Yun Xiang
She, Qunxin
An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title_full An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title_fullStr An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title_full_unstemmed An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title_short An archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
title_sort archaeal protein evolutionarily conserved in prokaryotes is a zinc-dependent metalloprotease
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497727/
https://www.ncbi.nlm.nih.gov/pubmed/22950735
http://dx.doi.org/10.1042/BSR20120074
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