Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1

Aims: To date, nine classes of copper regulation mechanisms have been discovered in bacteria and CsoR regulator protein is the most recent among them. Only a few have been structurally and functionally characterized. The present study was aimed to isolate and characterize the sequence, structure and...

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Main Authors: Musa, Nasihah, Mangavelu, Ashwaani, Au, Shaw Xian, Mohd Padzil, Azyyati, Kriznik, Alexandre, Mohammad Latif, Muhammad Alif, Jonet, Mohd Anuar, Ahmad Rodzli, Nazahiyah, Leow, Thean Chor, Mohamad Ali, Mohd Shukuri, Raja Abd Rahman, Raja Noor Zaliha, Salleh, Abu Bakar, M. Normi, Yahaya
Format: Article
Language:English
Published: Universiti Sains Malaysia 2024
Online Access:http://psasir.upm.edu.my/id/eprint/119836/
http://psasir.upm.edu.my/id/eprint/119836/1/119836.pdf
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author Musa, Nasihah
Mangavelu, Ashwaani
Au, Shaw Xian
Mohd Padzil, Azyyati
Kriznik, Alexandre
Mohammad Latif, Muhammad Alif
Jonet, Mohd Anuar
Ahmad Rodzli, Nazahiyah
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
Raja Abd Rahman, Raja Noor Zaliha
Salleh, Abu Bakar
M. Normi, Yahaya
author_facet Musa, Nasihah
Mangavelu, Ashwaani
Au, Shaw Xian
Mohd Padzil, Azyyati
Kriznik, Alexandre
Mohammad Latif, Muhammad Alif
Jonet, Mohd Anuar
Ahmad Rodzli, Nazahiyah
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
Raja Abd Rahman, Raja Noor Zaliha
Salleh, Abu Bakar
M. Normi, Yahaya
author_sort Musa, Nasihah
building UPM Institutional Repository
collection Online Access
description Aims: To date, nine classes of copper regulation mechanisms have been discovered in bacteria and CsoR regulator protein is the most recent among them. Only a few have been structurally and functionally characterized. The present study was aimed to isolate and characterize the sequence, structure and biochemical properties of CsoRGz-like hypothetical protein (HP) to be potentially used as a copper sensor protein. Methodology and results: A scan of the complete genome of a Geobacillus zalihae strain T1 revealed the presence of CsoR-like (CsoRGz) HP, which contains CsoR-like_DUF156 domain and highly conserved Cys-His-Cys residues essential for copper binding. It only shares moderate sequence identity with structurally characterized CsoR proteins. CsoRGz-like HP was subjected to sequence analyses to identify important domains, motifs and residues, while circular dichroism and X-ray crystallography were used to determine its secondary and tertiary structures. CsoRGz appears to be a dimer comprising mainly α-helices, with putative, conserved metal-binding ligands, Cys46-His71-Cys75, located on the α2 helix of the protein. Biophysical characterization of CsoRGz using UV/Vis and fluorescence spectrophotometry confirmed its interaction with Cu(I). Docking of Cu(I) to the dimeric structure of CsoRGz showed that Cu(I) could be coordinated by the above metal-binding residues, further stabilized by the hydrophobic core at the metal-binding site. Conclusion, significance and impact of study: The findings in this study suggest that CsoRGz-like HP might be a novel CsoR protein. This adds to the breadth and numbers of possible CsoR proteins, particularly uncharacterized ones, existing in the pool of hypothetical proteins, to be further characterized and compared across bacterial taxa.
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spelling upm-1198362025-09-19T08:14:04Z http://psasir.upm.edu.my/id/eprint/119836/ Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1 Musa, Nasihah Mangavelu, Ashwaani Au, Shaw Xian Mohd Padzil, Azyyati Kriznik, Alexandre Mohammad Latif, Muhammad Alif Jonet, Mohd Anuar Ahmad Rodzli, Nazahiyah Leow, Thean Chor Mohamad Ali, Mohd Shukuri Raja Abd Rahman, Raja Noor Zaliha Salleh, Abu Bakar M. Normi, Yahaya Aims: To date, nine classes of copper regulation mechanisms have been discovered in bacteria and CsoR regulator protein is the most recent among them. Only a few have been structurally and functionally characterized. The present study was aimed to isolate and characterize the sequence, structure and biochemical properties of CsoRGz-like hypothetical protein (HP) to be potentially used as a copper sensor protein. Methodology and results: A scan of the complete genome of a Geobacillus zalihae strain T1 revealed the presence of CsoR-like (CsoRGz) HP, which contains CsoR-like_DUF156 domain and highly conserved Cys-His-Cys residues essential for copper binding. It only shares moderate sequence identity with structurally characterized CsoR proteins. CsoRGz-like HP was subjected to sequence analyses to identify important domains, motifs and residues, while circular dichroism and X-ray crystallography were used to determine its secondary and tertiary structures. CsoRGz appears to be a dimer comprising mainly α-helices, with putative, conserved metal-binding ligands, Cys46-His71-Cys75, located on the α2 helix of the protein. Biophysical characterization of CsoRGz using UV/Vis and fluorescence spectrophotometry confirmed its interaction with Cu(I). Docking of Cu(I) to the dimeric structure of CsoRGz showed that Cu(I) could be coordinated by the above metal-binding residues, further stabilized by the hydrophobic core at the metal-binding site. Conclusion, significance and impact of study: The findings in this study suggest that CsoRGz-like HP might be a novel CsoR protein. This adds to the breadth and numbers of possible CsoR proteins, particularly uncharacterized ones, existing in the pool of hypothetical proteins, to be further characterized and compared across bacterial taxa. Universiti Sains Malaysia 2024 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/119836/1/119836.pdf Musa, Nasihah and Mangavelu, Ashwaani and Au, Shaw Xian and Mohd Padzil, Azyyati and Kriznik, Alexandre and Mohammad Latif, Muhammad Alif and Jonet, Mohd Anuar and Ahmad Rodzli, Nazahiyah and Leow, Thean Chor and Mohamad Ali, Mohd Shukuri and Raja Abd Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and M. Normi, Yahaya (2024) Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1. Malaysian Journal of Microbiology, 20 (4). pp. 417-429. ISSN 1823-8262; eISSN: 2231-7538 https://mjm.usm.my/index.php?r=cms/entry/view&id=2846&slug=Sequence%2C-structure-and-biophysical-characterization-of-CsoR-like-hypothetical-protein-from-Geobacillus-zalihae-strain-T1 10.21161/mjm.230265
spellingShingle Musa, Nasihah
Mangavelu, Ashwaani
Au, Shaw Xian
Mohd Padzil, Azyyati
Kriznik, Alexandre
Mohammad Latif, Muhammad Alif
Jonet, Mohd Anuar
Ahmad Rodzli, Nazahiyah
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
Raja Abd Rahman, Raja Noor Zaliha
Salleh, Abu Bakar
M. Normi, Yahaya
Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title_full Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title_fullStr Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title_full_unstemmed Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title_short Sequence, structure and biophysical characterization of CsoR-like hypothetical protein from Geobacillus zalihae strain T1
title_sort sequence, structure and biophysical characterization of csor-like hypothetical protein from geobacillus zalihae strain t1
url http://psasir.upm.edu.my/id/eprint/119836/
http://psasir.upm.edu.my/id/eprint/119836/
http://psasir.upm.edu.my/id/eprint/119836/
http://psasir.upm.edu.my/id/eprint/119836/1/119836.pdf