Bacterial cadherin domains as carbohydrate binding modules: Determination of affinity constants to insoluble complex polysaccharides

Milana Fraiberg, Ilya Borovok, Ronald M. Weiner, Raphael Lamed, Edward A. Bayer

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Cadherin (CA) and cadherin-like (CADG) doublet domains from the complex polysaccharide-degrading marine bacterium, Saccharophagus degradans 2-40, demonstrated reversible calcium-dependent binding to different complex polysaccharides, which serve as growth substrates for the bacterium. Here we describe a procedure based on adsorption of CA and CADG doublet domains to different insoluble complex polysaccharides, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for visualizing and quantifying the distribution of cadherins between the bound and unbound fractions. Scatchard plots were employed to determine the kinetics of interactions of CA and CADG with several complex carbohydrates. On the basis of these binding studies, the CA and CADG doublet domains are proposed to form a new family of carbohydrate-binding module (CBM).

Original languageEnglish
Title of host publicationBiomass Conversion
Subtitle of host publicationMethods and Protocols
EditorsMichael Himmel
Pages109-118
Number of pages10
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume908
ISSN (Print)1064-3745

Keywords

  • Binding affinity
  • CBM
  • Cadherin-like
  • Carbohydrate binding assay
  • SDS-PAGE
  • Scatchard plot

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