XIP-I, a xylanase inhibitor protein from wheat: A novel protein function

Nathalie Juge, Francoise Payan, Gary Williamson

Research output: Contribution to journalReview ArticleResearchpeer-review

91 Citations (Scopus)

Abstract

Endo-(1,4)-β-xylanases of plant and fungal origin play an important role in the degradation of arabinoxylans. Two distinct classes of proteinaceous endoxylanase inhibitors, the Triticum aestivum xylanase inhibitor (TAXI) and the xylanase inhibitor protein (XIP), have been identified in cereals. Engineering of proteins in conjunction with enzyme kinetics, thermodynamic, real-time interaction, and X-ray crystallographic studies has provided knowledge on the mechanism of inhibition of XIP-I towards endoxylanases. XIP-I is a 30 kDa protein which belongs to glycoside hydrolase family 18, and folds as a typical (β/α)8 barrel. Although the inhibitor shows highest homology with plant chitinases, XIP-I does not hydrolyse chitin; probably due to structural differences in the XIP-I binding cleft. The inhibitor is specific for fungal xylanases from glycoside hydrolases families 10 and 11, but does not inhibit bacterial enzymes. The inhibition is competitive and, depending on the xylanase, the Ki value can be as low as 3.4 nM. Site-directed mutagenesis of a xylanase from Aspergillus niger suggested that the XIP-I binding site was the conserved hairpin loop "thumb" region of family 11 xylanases. Furthermore, XIP-I shows the ability to inhibit barley α-amylases of glycoside hydrolase family 13, providing the first example of a protein able to inhibit members of different glycoside hydrolase families (10, 11, and 13), and additionally a novel function for a protein of glycoside hydrolase family 18.

Original languageEnglish
Pages (from-to)203-211
Number of pages9
JournalBBA Proteins and Proteomics
Volume1696
Issue number2
DOIs
Publication statusPublished - 12 Feb 2004
Externally publishedYes

Keywords

  • Barley α-amylase
  • Chitinase
  • Endo-(1,4)-β-xylanase
  • Protein-protein interaction
  • Site-directed mutagenesis
  • TAXI
  • Wheat proteinaceous endoxylanase inhibitor
  • X-ray structure
  • XIP

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