Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan

Abstract The global challenge of tuberculosis, caused by Mycobacterium tuberculosis (Mtb), is compounded by the emergence of drug-resistant strains. A critical factor in Mtb’s pathogenicity is its intricate cell envelope, which acts as a formidable barrier against immune defences and pharmacological...

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Main Authors: Yaqi Liu, Chelsea M. Brown, Satchal Erramilli, Yi-Chia Su, Shih-Yun Guu, Po-Sen Tseng, Yu-Jen Wang, Nam Ha Duong, Piotr Tokarz, Brian Kloss, Cheng-Ruei Han, Hung-Yu Chen, José Rodrigues, Kay-Hooi Khoo, Margarida Archer, Anthony A. Kossiakoff, Todd L. Lowary, Phillip J. Stansfeld, Rie Nygaard, Filippo Mancia
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58196-5
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author Yaqi Liu
Chelsea M. Brown
Satchal Erramilli
Yi-Chia Su
Shih-Yun Guu
Po-Sen Tseng
Yu-Jen Wang
Nam Ha Duong
Piotr Tokarz
Brian Kloss
Cheng-Ruei Han
Hung-Yu Chen
José Rodrigues
Kay-Hooi Khoo
Margarida Archer
Anthony A. Kossiakoff
Todd L. Lowary
Phillip J. Stansfeld
Rie Nygaard
Filippo Mancia
author_facet Yaqi Liu
Chelsea M. Brown
Satchal Erramilli
Yi-Chia Su
Shih-Yun Guu
Po-Sen Tseng
Yu-Jen Wang
Nam Ha Duong
Piotr Tokarz
Brian Kloss
Cheng-Ruei Han
Hung-Yu Chen
José Rodrigues
Kay-Hooi Khoo
Margarida Archer
Anthony A. Kossiakoff
Todd L. Lowary
Phillip J. Stansfeld
Rie Nygaard
Filippo Mancia
author_sort Yaqi Liu
collection DOAJ
description Abstract The global challenge of tuberculosis, caused by Mycobacterium tuberculosis (Mtb), is compounded by the emergence of drug-resistant strains. A critical factor in Mtb’s pathogenicity is its intricate cell envelope, which acts as a formidable barrier against immune defences and pharmacological interventions. Central to this envelope are arabinogalactan (AG) and lipoarabinomannan (LAM), two complex polysaccharides containing arabinan domains essential for maintaining cell wall structure and function. The arabinofuranosyltransferase AftB plays a pivotal role in the biosynthesis of these arabinan domains by catalyzing the addition of β-(1 → 2)-linked terminal arabinofuranose residues. Here, we present the cryo-EM structures of Mycobacterium chubuense AftB in both its apo form and bound to a donor substrate analog, resolved at 2.9 Å and 3.4 Å resolution, respectively. These structures reveal that AftB has a GT-C fold, with a transmembrane (TM) domain comprised of eleven TM helices and a periplasmic cap domain. AftB has a distinctive irregular, tube-shaped cavity that connects two proposed substrate binding sites. Through an integrated approach combining structural analysis, biochemical assays, and molecular dynamics simulations, we delineate the molecular basis of AftB’s reaction mechanism and propose a model for its catalytic function.
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spelling doaj-art-a11afe33b3a74eb89e054cb1adfd72402025-08-20T02:55:27ZengNature PortfolioNature Communications2041-17232025-04-0116111610.1038/s41467-025-58196-5Structural insights into terminal arabinosylation of mycobacterial cell wall arabinanYaqi Liu0Chelsea M. Brown1Satchal Erramilli2Yi-Chia Su3Shih-Yun Guu4Po-Sen Tseng5Yu-Jen Wang6Nam Ha Duong7Piotr Tokarz8Brian Kloss9Cheng-Ruei Han10Hung-Yu Chen11José Rodrigues12Kay-Hooi Khoo13Margarida Archer14Anthony A. Kossiakoff15Todd L. Lowary16Phillip J. Stansfeld17Rie Nygaard18Filippo Mancia19Department of Physiology and Cellular Biophysics, Columbia University Irving Medical CenterSchool of Life Sciences & Department of Chemistry, University of WarwickDepartment of Biochemistry and Molecular Biophysics, University of ChicagoInstitute of Biological Chemistry, Academia SinicaInstitute of Biological Chemistry, Academia SinicaDepartment of Chemistry, University of AlbertaInstitute of Biological Chemistry, Academia SinicaInstitute of Biological Chemistry, Academia SinicaDepartment of Biochemistry and Molecular Biophysics, University of ChicagoDepartment of Physiology and Cellular Biophysics, Columbia University Irving Medical CenterInstitute of Biological Chemistry, Academia SinicaInstitute of Biological Chemistry, Academia SinicaInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa (ITQB-UNL)Institute of Biological Chemistry, Academia SinicaInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa (ITQB-UNL)Department of Biochemistry and Molecular Biophysics, University of ChicagoInstitute of Biological Chemistry, Academia SinicaSchool of Life Sciences & Department of Chemistry, University of WarwickDepartment of Physiology and Cellular Biophysics, Columbia University Irving Medical CenterDepartment of Physiology and Cellular Biophysics, Columbia University Irving Medical CenterAbstract The global challenge of tuberculosis, caused by Mycobacterium tuberculosis (Mtb), is compounded by the emergence of drug-resistant strains. A critical factor in Mtb’s pathogenicity is its intricate cell envelope, which acts as a formidable barrier against immune defences and pharmacological interventions. Central to this envelope are arabinogalactan (AG) and lipoarabinomannan (LAM), two complex polysaccharides containing arabinan domains essential for maintaining cell wall structure and function. The arabinofuranosyltransferase AftB plays a pivotal role in the biosynthesis of these arabinan domains by catalyzing the addition of β-(1 → 2)-linked terminal arabinofuranose residues. Here, we present the cryo-EM structures of Mycobacterium chubuense AftB in both its apo form and bound to a donor substrate analog, resolved at 2.9 Å and 3.4 Å resolution, respectively. These structures reveal that AftB has a GT-C fold, with a transmembrane (TM) domain comprised of eleven TM helices and a periplasmic cap domain. AftB has a distinctive irregular, tube-shaped cavity that connects two proposed substrate binding sites. Through an integrated approach combining structural analysis, biochemical assays, and molecular dynamics simulations, we delineate the molecular basis of AftB’s reaction mechanism and propose a model for its catalytic function.https://doi.org/10.1038/s41467-025-58196-5
spellingShingle Yaqi Liu
Chelsea M. Brown
Satchal Erramilli
Yi-Chia Su
Shih-Yun Guu
Po-Sen Tseng
Yu-Jen Wang
Nam Ha Duong
Piotr Tokarz
Brian Kloss
Cheng-Ruei Han
Hung-Yu Chen
José Rodrigues
Kay-Hooi Khoo
Margarida Archer
Anthony A. Kossiakoff
Todd L. Lowary
Phillip J. Stansfeld
Rie Nygaard
Filippo Mancia
Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
Nature Communications
title Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
title_full Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
title_fullStr Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
title_full_unstemmed Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
title_short Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
title_sort structural insights into terminal arabinosylation of mycobacterial cell wall arabinan
url https://doi.org/10.1038/s41467-025-58196-5
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