Structure of the priming arabinosyltransferase AftA required for AG biosynthesis of <i>Mycobacterium tuberculosis</i>.

Abstract

Arabinogalactan (AG) is an essential cell wall component in mycobacterial species, including the deadly human pathogen <i>Mycobacterium tuberculosis</i>. It plays a pivotal role in forming the rigid mycolyl-AG-peptidoglycan core for in vitro growth. AftA is a membrane-bound arabinosyltransferase and a key enzyme involved in AG biosynthesis which bridges the assembly of the arabinan chain to the galactan chain. It is known that AftA catalyzes the transfer of the first arabinofuranosyl residue from the donor decaprenyl-monophosphoryl-arabinose to the mature galactan chain (i.e., priming); however, the priming mechanism remains elusive. Herein, we report the cryo-EM structure of <i>Mtb</i> AftA. The detergent-embedded AftA assembles as a dimer with an interface maintained by both the transmembrane domain (TMD) and the soluble C-terminal domain (CTD) in the periplasm. The structure shows a conserved glycosyltransferase-C fold and two cavities converging at the active site. A metal ion participates in the interaction of TMD and CTD of each AftA molecule. Structural analyses combined with functional mutagenesis suggests a priming mechanism catalyzed by AftA in <i>Mtb</i> AG biosynthesis. Our data further provide a unique perspective into anti-TB drug discovery.