EMD-21600
Single-Particle Cryo-EM Structure of Arabinofuranosyltransferase AftD from Mycobacteria, Mutant R1389S Class 1
EMD-21600
Single-particle3.5 Å
Deposition: 27/03/2020Map released: 13/05/2020
Last modified: 13/11/2024
Sample Organism:
Mycobacteroides abscessus
Sample: Mutant R1389S Class 1 Mycobacterial Arabinofuranosyltransferase AftD Complexed with Acyl Carrier Protein
Fitted models: 6wbx (Avg. Q-score: 0.518)
Raw data: EMPIAR-10391
Deposition Authors: Tan YZ
,
Zhang L
Sample: Mutant R1389S Class 1 Mycobacterial Arabinofuranosyltransferase AftD Complexed with Acyl Carrier Protein
Fitted models: 6wbx (Avg. Q-score: 0.518)
Raw data: EMPIAR-10391
Deposition Authors: Tan YZ
,
Zhang L
Cryo-EM Structures and Regulation of Arabinofuranosyltransferase AftD from Mycobacteria.
Tan YZ ,
Zhang L ,
Rodrigues J ,
Zheng RB,
Giacometti SI ,
Rosario AL,
Kloss B,
Dandey VP,
Wei H,
Brunton R,
Raczkowski AM ,
Athayde D ,
Catalao MJ ,
Pimentel M,
Clarke OB,
Lowary TL,
Archer M ,
Niederweis M ,
Potter CS,
Carragher B,
Mancia F
(2020) Mol Cell , 78 , 683
,
Zhang L ,
Rodrigues J ,
Zheng RB,
Giacometti SI ,
Rosario AL,
Kloss B,
Dandey VP,
Wei H,
Brunton R,
Raczkowski AM ,
Athayde D ,
Catalao MJ ,
Pimentel M,
Clarke OB,
Lowary TL,
Archer M ,
Niederweis M ,
Potter CS,
Carragher B,
Mancia F
(2020) Mol Cell , 78 , 683
Abstract:
Mycobacterium tuberculosis causes tuberculosis, a disease that kills over 1 million people each year. Its cell envelope is a common antibiotic target and has a unique structure due, in part, to two lipidated polysaccharides-arabinogalactan and lipoarabinomannan. Arabinofuranosyltransferase D (AftD) is an essential enzyme involved in assembling these glycolipids. We present the 2.9-Å resolution structure of M. abscessus AftD, determined by single-particle cryo-electron microscopy. AftD has a conserved GT-C glycosyltransferase fold and three carbohydrate-binding modules. Glycan array analysis shows that AftD binds complex arabinose glycans. Additionally, AftD is non-covalently complexed with an acyl carrier protein (ACP). 3.4- and 3.5-Å structures of a mutant with impaired ACP binding reveal a conformational change, suggesting that ACP may regulate AftD function. Mutagenesis experiments using a conditional knockout constructed in M. smegmatis confirm the essentiality of the putative active site and the ACP binding for AftD function.
Mycobacterium tuberculosis causes tuberculosis, a disease that kills over 1 million people each year. Its cell envelope is a common antibiotic target and has a unique structure due, in part, to two lipidated polysaccharides-arabinogalactan and lipoarabinomannan. Arabinofuranosyltransferase D (AftD) is an essential enzyme involved in assembling these glycolipids. We present the 2.9-Å resolution structure of M. abscessus AftD, determined by single-particle cryo-electron microscopy. AftD has a conserved GT-C glycosyltransferase fold and three carbohydrate-binding modules. Glycan array analysis shows that AftD binds complex arabinose glycans. Additionally, AftD is non-covalently complexed with an acyl carrier protein (ACP). 3.4- and 3.5-Å structures of a mutant with impaired ACP binding reveal a conformational change, suggesting that ACP may regulate AftD function. Mutagenesis experiments using a conditional knockout constructed in M. smegmatis confirm the essentiality of the putative active site and the ACP binding for AftD function.