Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter

Spellmon, Nicholas; Muszyński, Artur; Górniak, Ireneusz; Vlach, Jiri; Hahn, David; Azadi, Parastoo; Zimmer, Jochen

doi:10.1038/s41467-022-32597-2

Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter

Journal Article · Mon Sep 05 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-32597-2· OSTI ID:1962865

^[1]; ^[2]; ^[3]; ^[2]; Hahn, David ^[2]; Azadi, Parastoo ^[2]; ^[3]

Univ. of Virginia School of Medicine, Charlottesville, VA (United States); University of Virginia
Univ. of Georgia, Athens, GA (United States)
Univ. of Virginia School of Medicine, Charlottesville, VA (United States)

O antigens are ubiquitous protective extensions of lipopolysaccharides in the extracellular leaflet of the Gram-negative outer membrane. Following biosynthesis in the cytosol, the lipid-linked polysaccharide is transported to the periplasm by the WzmWzt ABC transporter. Often, O antigen secretion requires the chemical modification of its elongating terminus, which the transporter recognizes via a carbohydrate-binding domain (CBD). Here, using components from A. aeolicus, we identify the O antigen structure with methylated mannose or rhamnose as its cap. Crystal and cryo electron microscopy structures reveal how WzmWzt recognizes this cap between its carbohydrate and nucleotide-binding domains in a nucleotide-free state. ATP binding induces drastic conformational changes of its CBD, terminating interactions with the O antigen. ATPase assays and site directed mutagenesis reveal reduced hydrolytic activity upon O antigen binding, likely to facilitate polymer loading into the ABC transporter. Our results elucidate critical steps in the recognition and translocation of polysaccharides by ABC transporters.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center

Sponsoring Organization:: National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: SC0015662

OSTI ID:: 1962865

Alternate ID(s):: OSTI ID: 1900964

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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