Dataset Information

Alternating L4 loop architecture of the bacterial polysaccharide co-polymerase WzzE.

ABSTRACT: Lipopolysaccharides such as the enterobacterial common antigen are important components of the enterobacterial cell envelope that act as a protective barrier against the environment and are often polymerized by the inner membrane bound Wzy-dependent pathway. By employing cryo-electron microscopy we show that WzzE, the co-polymerase component of this pathway that is responsible for the length modulation of the enterobacterial common antigen, is octameric with alternating up-down conformations of its L4 loops. The alternating up-down nature of these essential loops, located at the top of the periplasmic bell, are modulated by clashing helical faces between adjacent protomers that flank the L4 loops around the octameric periplasmic bell. This alternating arrangement and a highly negatively charged binding face create a dynamic environment in which the polysaccharide chain is extended, and suggest a ratchet-type mechanism for polysaccharide elongation.

SUBMITTER: Wiseman B

PROVIDER: S-EPMC10397196 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Alternating L4 loop architecture of the bacterial polysaccharide co-polymerase WzzE.

Wiseman Benjamin B Widmalm Göran G Högbom Martin M

Communications biology 20230802 1

Lipopolysaccharides such as the enterobacterial common antigen are important components of the enterobacterial cell envelope that act as a protective barrier against the environment and are often polymerized by the inner membrane bound Wzy-dependent pathway. By employing cryo-electron microscopy we show that WzzE, the co-polymerase component of this pathway that is responsible for the length modulation of the enterobacterial common antigen, is octameric with alternating up-down conformations of ...[more]

PMID: 37532793

Similar Datasets

Project description:The ability of bacteria to synthesise complex polysaccharide chains at a controlled number of repeating units has wide implications for a range of biological activities that include: symbiosis, biofilm formation and immune system avoidance. Complex polysaccharide chains such as the O antigen (Oag) component of lipopolysaccharide and the enterobacterial common antigen (ECA) are synthesised by the most common polysaccharide synthesis pathway used in bacteria, known as the Wzy-dependent pathway. The Oag and ECA are polymerized into chains via the inner membrane proteins WzyB and WzyE, respectively, while the respective co-polymerases WzzB and WzzE modulate the number of repeat units in the chains or "the modal length" of the polysaccharide via a hypothesised interaction. Our data shows for the first time "cross-talk" between Oag and ECA synthesis in that WzzE is able to partially regulate Oag modal length via a potential interaction with WzyB. To investigate this, one or both of the transmembrane regions (TM1 and TM2) of WzzE and WzzB were swapped creating six chimera proteins. Several chimeric proteins showed significant increases Oag modal length control, while others reduced control. Additionally, co-purification experiments show an interaction between WzyB and WzzB for the first time without the use of a chemical cross-linker, and a novel interaction between WzyB and WzzE. These results suggest the TM2 region of Wzz proteins plays a critical role in Oag and ECA modal length control, presumably via the interaction with respective Wzy proteins, thus providing insight into the complex mechanism underlying the control of polysaccharide biosynthesis.ImportanceBacteria synthesise complex polysaccharide chains at a controlled number of repeating units, this has wide implications for a range of bacterial activities involved in virulence. Examples of complex polysaccharide chains include, the O antigen (Oag) component of lipopolysaccharide and the enterobacterial common antigen (ECA), both of these examples are predominantly synthesised by their own independent Wzy-dependent pathway. Our data show for the first time "cross-talk" between Oag and ECA synthesis and identifies novel physical protein-protein interactions between proteins in these systems. These findings further the understanding of how the system functions to control polysaccharide chain length which has great implications for novel biotechnologies and/or the combat of bacterial diseases.

Dataset Information

Alternating L4 loop architecture of the bacterial polysaccharide co-polymerase WzzE.

Publications

Alternating L4 loop architecture of the bacterial polysaccharide co-polymerase WzzE.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets