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Related Concept Videos

Secondary Active Transport01:55

Secondary Active Transport

One example of how cells use the energy contained in electrochemical gradients is demonstrated by glucose transport into cells. The ion vital to this process is sodium (Na+), which is typically present in higher concentrations extracellularly than in the cytosol. Such a concentration difference is due, in part, to the action of an enzyme “pump” embedded in the cellular membrane that actively expels Na+ from a cell. Importantly, as this pump contributes to the high concentration of...
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ATP-binding cassette or ABC transporter is the largest superfamily of integral membrane proteins. The transporters have transmembrane-binding domains (TMDs) and nucleotide-binding domains (NBDs). The TMDs are specific to their substrates, whereas the NBDs are similar to engines that complete ATP hydrolysis to complete the substrate transport. They can be full transporters consisting of two TMDs and NBDs, half transporters with one TMD and NBD, while some encoded with a single TMD or NBD are...
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ATP-binding cassette or ABC transporters are a class of ATP-driven pumps that hydrolyze ATP to move solutes across the membrane. They can be grouped into importers and exporters. While exporters are present in all domains of life, importers exist only in bacteria and some plants.
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Substrate binding by a bacterial ABC transporter involved in polysaccharide export.

Leslie Cuthbertson1, Matthew S Kimber, Chris Whitfield

  • 1Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.

Proceedings of the National Academy of Sciences of the United States of America
|November 23, 2007
PubMed
Summary

ATP-binding-cassette (ABC) transporters export bacterial glycoconjugates. The study reveals a unique C-terminal domain in Wzt that specifically binds O-antigenic polysaccharide (O-PS), crucial for Gram-negative pathogen virulence.

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Area of Science:

  • Microbiology
  • Structural Biology
  • Biochemistry

Background:

  • ATP-binding-cassette (ABC) transporters facilitate the export of diverse cell-surface glycoconjugates in bacteria.
  • Lipopolysaccharide (LPS) O-antigenic polysaccharide (O-PS) is a key virulence factor in Gram-negative pathogens, exported by ABC transporters.
  • Two distinct pathways exist for O-PS synthesis, with Escherichia coli O8 and O9a serving as model systems for ABC transporter-mediated export.

Purpose of the Study:

  • To investigate the mechanism of O-PS export by ABC transporters, focusing on the role of the Wzt nucleotide-binding component.
  • To elucidate the structure and function of the unique C-terminal domain of Wzt (C-Wzt) in O-PS recognition and specificity.
  • To understand how O-PS export is coordinated with its biosynthesis and termination.

Main Methods:

  • Expression and functional analysis of Wzt domains as separate polypeptides.
  • In vitro binding assays to determine C-Wzt's interaction with O-PS.
  • X-ray crystallography to determine the structure of C-Wzt(O9a).

Main Results:

  • Both N-terminal and C-terminal domains of Wzt are essential for O-PS export, and can function independently.
  • The C-terminal domain (C-Wzt) specifically binds O-PS at its nonreducing end.
  • The crystal structure of C-Wzt(O9a) reveals an immunoglobulin-like beta-sandwich fold containing the O-PS binding pocket.

Conclusions:

  • The C-terminal domain of Wzt represents a novel mechanism for substrate recognition in ABC transporters.
  • This finding provides critical insights into the coordinated regulation of O-PS biosynthesis, termination, and export.
  • Understanding this system is vital for targeting virulence mechanisms in Gram-negative pathogens.