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Title | Cryo-EM Structures and Regulation of Arabinofuranosyltransferase AftD from Mycobacteria. |
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Journal, issue, pages | Mol Cell, Vol. 78, Issue 4, Page 683-699.e11, Year 2020 |
Publish date | May 21, 2020 |
Authors | Yong Zi Tan / Lei Zhang / José Rodrigues / Ruixiang Blake Zheng / Sabrina I Giacometti / Ana L Rosário / Brian Kloss / Venkata P Dandey / Hui Wei / Richard Brunton / Ashleigh M Raczkowski / Diogo Athayde / Maria João Catalão / Madalena Pimentel / Oliver B Clarke / Todd L Lowary / Margarida Archer / Michael Niederweis / Clinton S Potter / Bridget Carragher / Filippo Mancia / |
PubMed 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 ...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. |
External links | Mol Cell / PubMed:32386575 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.9 - 3.5 Å |
Structure data | EMDB-21580, PDB-6w98: EMDB-21600, PDB-6wbx: EMDB-21601, PDB-6wby: |
Chemicals | ChemComp-CA: ChemComp-6OU: ChemComp-PNS: ChemComp-HOH: |
Source |
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Keywords | MEMBRANE PROTEIN / Glycosyltransferase / lipomannan / lipoarabinomannan / arabinofuranose / nanodisc / single-particle cryo-electron microscopy / acyl carrier protein |