Plant Molecular Biology

, Volume 71, Issue 3, pp 277–289 | Cite as

The first crystal structures of a family 19 class IV chitinase: the enzyme from Norway spruce

  • Wimal Ubhayasekera
  • Reetika Rawat
  • Sharon Wing Tak Ho
  • Malgorzata Wiweger
  • Sara Von Arnold
  • Mee-Len Chye
  • Sherry L. Mowbray


Chitinases help plants defend themselves against fungal attack, and play roles in other processes, including development. The catalytic modules of most plant chitinases belong to glycoside hydrolase family 19. We report here x-ray structures of such a module from a Norway spruce enzyme, the first for any family 19 class IV chitinase. The bi-lobed structure has a wide cleft lined by conserved residues; the most interesting for catalysis are Glu113, the proton donor, and Glu122, believed to be a general base that activate a catalytic water molecule. Comparisons to class I and II enzymes show that loop deletions in the class IV proteins make the catalytic cleft shorter and wider; from modeling studies, it is predicted that only three N-acetylglucosamine-binding subsites exist in class IV. Further, the structural comparisons suggest that the family 19 enzymes become more closed on substrate binding. Attempts to solve the structure of the complete protein including the associated chitin-binding module failed, however, modeling studies based on close relatives indicate that the binding module recognizes at most three N-acetylglucosamine units. The combined results suggest that the class IV enzymes are optimized for shorter substrates than the class I and II enzymes, or alternatively, that they are better suited for action on substrates where only small regions of chitin chain are accessible. Intact spruce chitinase is shown to possess antifungal activity, which requires the binding module; removing this module had no effect on measured chitinase activity.


Chitinase Family 19 Picea abies Norway spruce Conformational changes Class IV 



The authors would like to thank Dr. Fred Asiegbu (Swedish University of Agricultural Sciences) for providing Heterobasidion annosum (strain FP5), and Dr. Mark Harris (Uppsala University) for photographing the chitinase-inhibited fungal plate. The work was supported by the Swedish Research Council (VR) and the Swedish Foundation for Strategic Research via the Glycoconjugates in Biological Systems network, GLIBS (SLM), as well as by the University of Hong Kong (ORA10208034) (MLC).


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Wimal Ubhayasekera
    • 1
  • Reetika Rawat
    • 2
  • Sharon Wing Tak Ho
    • 2
  • Malgorzata Wiweger
    • 3
  • Sara Von Arnold
    • 4
  • Mee-Len Chye
    • 2
  • Sherry L. Mowbray
    • 1
  1. 1.Department of Molecular Biology, Biomedical CenterSwedish University of Agricultural SciencesUppsalaSweden
  2. 2.School of Biological SciencesThe University of Hong KongHong KongChina
  3. 3.Department of PathologyLeiden University Medical CenterLeidenThe Netherlands
  4. 4.Department of Plant Biology and Forest GeneticsSwedish University of Agricultural SciencesUppsalaSweden

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