Search for a dextransucrase minimal motif involved in dextran binding

FEBS Lett. 2007 Oct 2;581(24):4675-80. doi: 10.1016/j.febslet.2007.08.062. Epub 2007 Sep 4.

Abstract

Fourteen truncated forms of Leuconostoc mesenteroides NRRL B512-F dextransucrase, involving N-, C- or N- plus C-terminal domain truncations were tested for their ability to bind dextrans. The shortest fragment (14kDa molecular weight) that still exhibited a strong interaction with dextran was localized between amino acids N1397 and A1527 of the C-terminal domain (GBD-7) and consists of six YG repeats. With a dissociation constant K(d) of 2.8x10(-9)M, this motif shows a very high affinity for isomaltohexaose and longer dextrans, supporting the proposed role of GBD in polymer formation. The potential application of GBD-7 as an affinity tag onto cheap resins like Sephacryl S300HR for rapid purification was evaluated and is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Dextrans / metabolism*
  • Gene Deletion
  • Glucans / metabolism
  • Glucosyltransferases / chemistry
  • Glucosyltransferases / genetics
  • Glucosyltransferases / isolation & purification
  • Glucosyltransferases / metabolism*
  • Leuconostoc / genetics
  • Leuconostoc / metabolism
  • Molecular Sequence Data
  • Protein Binding
  • Sequence Alignment
  • Thioredoxins / genetics
  • Thioredoxins / isolation & purification
  • Thioredoxins / metabolism

Substances

  • Dextrans
  • Glucans
  • Thioredoxins
  • Glucosyltransferases
  • dextransucrase