Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>

<i>Background and Objective</i>: ATP-binding cassette (ABC) transporters are prominent drug targets due to their highly efficient trafficking capabilities and their significant physiological and clinical roles. Gaining insight into their biophysical and biomechanistic properties is cruci...

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Main Authors: Aisha Farhana, Abdullah Alsrhani, Hasan Ejaz, Muharib Alruwaili, Ayman A. M. Alameen, Emad Manni, Zafar Rasheed, Yusuf Saleem Khan
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Medicina
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Online Access:https://www.mdpi.com/1648-9144/60/11/1891
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author Aisha Farhana
Abdullah Alsrhani
Hasan Ejaz
Muharib Alruwaili
Ayman A. M. Alameen
Emad Manni
Zafar Rasheed
Yusuf Saleem Khan
author_facet Aisha Farhana
Abdullah Alsrhani
Hasan Ejaz
Muharib Alruwaili
Ayman A. M. Alameen
Emad Manni
Zafar Rasheed
Yusuf Saleem Khan
author_sort Aisha Farhana
collection DOAJ
description <i>Background and Objective</i>: ATP-binding cassette (ABC) transporters are prominent drug targets due to their highly efficient trafficking capabilities and their significant physiological and clinical roles. Gaining insight into their biophysical and biomechanistic properties is crucial to maximize their pharmacological potential. <i>Materials and Methods</i>: In this study, we present the biochemical and biophysical characterization, and phylogenetic analysis of the domains of <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>) ABC transporters: the exporter Rv1348 (IrtA) and the importer system Rv1349-Rv2895c (IrtB-Rv2895c), both involved in siderophore-mediated iron uptake. <i>Results</i>: Our findings reveal that the substrate-binding domain (SBD) of IrtA functions as an active monomer, while Rv2895c, which facilitates the uptake of siderophore-bound iron, exists in a dynamic equilibrium between dimeric and monomeric forms. Furthermore, ATP binding induces the dimerization of the ATPase domains in both IrtA (ATPase I) and IrtB (ATPaseII), but only the ATPase domain of IrtA (ATPase I) is active independently. We also analyzed the stability of substrate binding to the domains of the two transporters across varying temperature and pH ranges, revealing significant shifts in their activity under different conditions. Our study highlights the conformational changes that accompany substrate interaction with the transporter domains, providing insights into the fundamental mechanism required for the translocation of siderophore to the extracytoplasmic milieu by IrtB and, subsequently, import of their ferrated forms by the IrtB-Rv2895c complex. Phylogenetic analyses based on ATPase domains reveal that IrtA shares features with both archaeal and eukaryotic transporters, while IrtB is unique to mycobacterial species. <i>Conclusions</i>: Together, these findings provide valuable insights, which could accelerate the development of intervention strategies for this critical pathway pivotal in the progression of <i>M. tuberculosis</i> infection.
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spelling doaj-art-f4edcc69a5594bf988204786b5a815fb2025-08-20T02:04:59ZengMDPI AGMedicina1010-660X1648-91442024-11-016011189110.3390/medicina60111891Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>Aisha Farhana0Abdullah Alsrhani1Hasan Ejaz2Muharib Alruwaili3Ayman A. M. Alameen4Emad Manni5Zafar Rasheed6Yusuf Saleem Khan7Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Aljouf, Saudi ArabiaDepartment of Pathology, College of Medicine, Qassim University, Buraidah 51452, Qassim, Saudi ArabiaDepartment of Anatomy, College of Medicine, University of Hail, Hail 55476, Hail, Saudi Arabia<i>Background and Objective</i>: ATP-binding cassette (ABC) transporters are prominent drug targets due to their highly efficient trafficking capabilities and their significant physiological and clinical roles. Gaining insight into their biophysical and biomechanistic properties is crucial to maximize their pharmacological potential. <i>Materials and Methods</i>: In this study, we present the biochemical and biophysical characterization, and phylogenetic analysis of the domains of <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>) ABC transporters: the exporter Rv1348 (IrtA) and the importer system Rv1349-Rv2895c (IrtB-Rv2895c), both involved in siderophore-mediated iron uptake. <i>Results</i>: Our findings reveal that the substrate-binding domain (SBD) of IrtA functions as an active monomer, while Rv2895c, which facilitates the uptake of siderophore-bound iron, exists in a dynamic equilibrium between dimeric and monomeric forms. Furthermore, ATP binding induces the dimerization of the ATPase domains in both IrtA (ATPase I) and IrtB (ATPaseII), but only the ATPase domain of IrtA (ATPase I) is active independently. We also analyzed the stability of substrate binding to the domains of the two transporters across varying temperature and pH ranges, revealing significant shifts in their activity under different conditions. Our study highlights the conformational changes that accompany substrate interaction with the transporter domains, providing insights into the fundamental mechanism required for the translocation of siderophore to the extracytoplasmic milieu by IrtB and, subsequently, import of their ferrated forms by the IrtB-Rv2895c complex. Phylogenetic analyses based on ATPase domains reveal that IrtA shares features with both archaeal and eukaryotic transporters, while IrtB is unique to mycobacterial species. <i>Conclusions</i>: Together, these findings provide valuable insights, which could accelerate the development of intervention strategies for this critical pathway pivotal in the progression of <i>M. tuberculosis</i> infection.https://www.mdpi.com/1648-9144/60/11/1891<i>Mycobacterium tuberculosis</i>siderophore-iron uptakeABC transportersRv1348 (IrtA) and Rv2895c (IrtB)evolutionary linkage of transportersdrug targeting
spellingShingle Aisha Farhana
Abdullah Alsrhani
Hasan Ejaz
Muharib Alruwaili
Ayman A. M. Alameen
Emad Manni
Zafar Rasheed
Yusuf Saleem Khan
Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
Medicina
<i>Mycobacterium tuberculosis</i>
siderophore-iron uptake
ABC transporters
Rv1348 (IrtA) and Rv2895c (IrtB)
evolutionary linkage of transporters
drug targeting
title Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
title_full Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
title_fullStr Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
title_full_unstemmed Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
title_short Substrate-Induced Structural Dynamics and Evolutionary Linkage of Siderophore-Iron ABC Transporters of <i>Mycobacterium tuberculosis</i>
title_sort substrate induced structural dynamics and evolutionary linkage of siderophore iron abc transporters of i mycobacterium tuberculosis i
topic <i>Mycobacterium tuberculosis</i>
siderophore-iron uptake
ABC transporters
Rv1348 (IrtA) and Rv2895c (IrtB)
evolutionary linkage of transporters
drug targeting
url https://www.mdpi.com/1648-9144/60/11/1891
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