7K2T

Mg2+/ATP-bound structure of the full-length WzmWzt O antigen ABC transporter in lipid nanodiscs

Summary for 7K2T

• Entry DOI: 10.2210/pdb7k2t/pdb
• EMDB information: 22644
• Descriptor: ABC transporter, Transport permease protein, ADENOSINE-5'-TRIPHOSPHATE, ... (4 entities in total)
• Functional Keywords: o antigen transporter, integral membrane protein, lipopolysaccharide lps biosynthesis, transport protein
• Biological source: Aquifex aeolicus (strain VF5); More
• Total number of polymer chains: 4
• Total formula weight: 153685.57

Authors

Caffalette, C.A.,Zimmer, J. (deposition date: 2020-09-09, release date: 2021-01-13, Last modification date: 2024-03-06)

Primary citation

Caffalette, C.A.,Zimmer, J.
Cryo-EM structure of the full-length WzmWzt ABC transporter required for lipid-linked O antigen transport.
Proc.Natl.Acad.Sci.USA, 118:-, 2021

PubMed Abstract: O antigens are important cell surface polysaccharides in gram-negative bacteria where they extend core lipopolysaccharides in the extracellular leaflet of the outer membrane. O antigen structures are serotype specific and form extended cell surface barriers endowing many pathogens with survival benefits. In the ABC transporter-dependent biosynthesis pathway, O antigens are assembled on the cytosolic side of the inner membrane on a lipid anchor and reoriented to the periplasmic leaflet by the channel-forming WzmWzt ABC transporter for ligation to the core lipopolysaccharides. In many cases, this process depends on the chemical modification of the O antigen's nonreducing terminus, sensed by WzmWzt via a carbohydrate-binding domain (CBD) that extends its nucleotide-binding domain (NBD). Here, we provide the cryo-electron microscopy structure of the full-length WzmWzt transporter from bound to adenosine triphosphate (ATP) and in a lipid environment, revealing a highly asymmetric transporter organization. The CBDs dimerize and associate with only one NBD. Conserved loops at the CBD dimer interface straddle a conserved peripheral NBD helix. The CBD dimer is oriented perpendicularly to the NBDs and its putative ligand-binding sites face the transporter to likely modulate ATPase activity upon O antigen binding. Further, our structure reveals a closed WzmWzt conformation in which an aromatic belt near the periplasmic channel exit seals the transporter in a resting, ATP-bound state. The sealed transmembrane channel is asymmetric, with one open and one closed cytosolic and periplasmic portal. The structure provides important insights into O antigen recruitment to and translocation by WzmWzt and related ABC transporters.

PubMed: 33443152
DOI: 10.1073/pnas.2016144118

Experimental method

ELECTRON MICROSCOPY (3.6 Å)