A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library
The presented study examines the phenomenon of the fluorescence under UV light excitation (312 nm) of E. coli cells expressing a novel metagenomic-derived putative methylthioadenosine phosphorylase gene, called rsfp, grown on LB agar supplemented with a fluorescent dye rhodamine B. For this purpose,...
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ftpubmed:oai:pubmedcentral.nih.gov:3561333 2023-05-15T13:54:18+02:00 A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library Bartasun, Paulina Cieśliński, Hubert Bujacz, Anna Wierzbicka-Woś, Anna Kur, Józef 2013-01-31 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561333 http://www.ncbi.nlm.nih.gov/pubmed/23383268 https://doi.org/10.1371/journal.pone.0055697 en eng Public Library of Science http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561333 http://www.ncbi.nlm.nih.gov/pubmed/23383268 http://dx.doi.org/10.1371/journal.pone.0055697 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 credited. CC-BY Research Article Text 2013 ftpubmed https://doi.org/10.1371/journal.pone.0055697 2013-09-04T19:17:52Z The presented study examines the phenomenon of the fluorescence under UV light excitation (312 nm) of E. coli cells expressing a novel metagenomic-derived putative methylthioadenosine phosphorylase gene, called rsfp, grown on LB agar supplemented with a fluorescent dye rhodamine B. For this purpose, an rsfp gene was cloned and expressed in an LMG194 E. coli strain using an arabinose promoter. The resulting RSFP protein was purified and its UV-VIS absorbance spectrum and emission spectrum were assayed. Simultaneously, the same spectroscopic studies were carried out for rhodamine B in the absence or presence of RSFP protein or native E. coli proteins, respectively. The results of the spectroscopic studies suggested that the fluorescence of E. coli cells expressing rsfp gene under UV illumination is due to the interaction of rhodamine B molecules with the RSFP protein. Finally, this interaction was proved by a crystallographic study and then by site-directed mutagenesis of rsfp gene sequence. The crystal structures of RSFP apo form (1.98 Å) and complex RSFP/RB (1.90 Å) show a trimer of RSFP molecules located on the crystallographic six fold screw axis. The RSFP complex with rhodamine B revealed the binding site for RB, in the pocket located on the interface between symmetry related monomers. Text Antarc* Antarctic PubMed Central (PMC) Antarctic PLoS ONE 8 1 e55697 |
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Research Article Bartasun, Paulina Cieśliński, Hubert Bujacz, Anna Wierzbicka-Woś, Anna Kur, Józef A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
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Research Article |
description |
The presented study examines the phenomenon of the fluorescence under UV light excitation (312 nm) of E. coli cells expressing a novel metagenomic-derived putative methylthioadenosine phosphorylase gene, called rsfp, grown on LB agar supplemented with a fluorescent dye rhodamine B. For this purpose, an rsfp gene was cloned and expressed in an LMG194 E. coli strain using an arabinose promoter. The resulting RSFP protein was purified and its UV-VIS absorbance spectrum and emission spectrum were assayed. Simultaneously, the same spectroscopic studies were carried out for rhodamine B in the absence or presence of RSFP protein or native E. coli proteins, respectively. The results of the spectroscopic studies suggested that the fluorescence of E. coli cells expressing rsfp gene under UV illumination is due to the interaction of rhodamine B molecules with the RSFP protein. Finally, this interaction was proved by a crystallographic study and then by site-directed mutagenesis of rsfp gene sequence. The crystal structures of RSFP apo form (1.98 Å) and complex RSFP/RB (1.90 Å) show a trimer of RSFP molecules located on the crystallographic six fold screw axis. The RSFP complex with rhodamine B revealed the binding site for RB, in the pocket located on the interface between symmetry related monomers. |
format |
Text |
author |
Bartasun, Paulina Cieśliński, Hubert Bujacz, Anna Wierzbicka-Woś, Anna Kur, Józef |
author_facet |
Bartasun, Paulina Cieśliński, Hubert Bujacz, Anna Wierzbicka-Woś, Anna Kur, Józef |
author_sort |
Bartasun, Paulina |
title |
A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
title_short |
A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
title_full |
A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
title_fullStr |
A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
title_full_unstemmed |
A Study on the Interaction of Rhodamine B with Methylthioadenosine Phosphorylase Protein Sourced from an Antarctic Soil Metagenomic Library |
title_sort |
study on the interaction of rhodamine b with methylthioadenosine phosphorylase protein sourced from an antarctic soil metagenomic library |
publisher |
Public Library of Science |
publishDate |
2013 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561333 http://www.ncbi.nlm.nih.gov/pubmed/23383268 https://doi.org/10.1371/journal.pone.0055697 |
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Antarctic |
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Antarctic |
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Antarc* Antarctic |
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Antarc* Antarctic |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561333 http://www.ncbi.nlm.nih.gov/pubmed/23383268 http://dx.doi.org/10.1371/journal.pone.0055697 |
op_rights |
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 credited. |
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CC-BY |
op_doi |
https://doi.org/10.1371/journal.pone.0055697 |
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PLoS ONE |
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e55697 |
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