This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Derda, M.
Right arrow Articles by Linder, E.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Derda, M.
Right arrow Articles by Linder, E.
Agricola
Right arrow Articles by Derda, M.
Right arrow Articles by Linder, E.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2009, p. 6827-6830, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.01555-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Labeled Trichoderma reesei Cellulase as a Marker for Acanthamoeba Cyst Wall Cellulose in Infected Tissues {triangledown}

Monika Derda,1* Jadwiga Winiecka-Krusnell,2 Markus B. Linder,3 and Ewert Linder2,4

Department of Biology and Medical Parasitology, University of Medical Sciences, 61-701 Poznan, Poland,1 Swedish Institute for Infectious Disease Control, 171 82 Solna, Sweden,2 VTT Biotechnology, VTT Technical Research Centre of Finland, FIN 02044-VTT, Espoo, Finland,3 Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 82 Solna, Sweden4

Received 2 July 2009/ Accepted 28 August 2009

Some protozoans are able to encyst as a protective response to a harmful environment. The cyst wall usually contains chitin as its main structural constituent. Acanthamoeba is an exception since its cyst wall contains cellulose. Specific cytochemical differentiation between cellulose and chitin by microscopy has not been possible due to the similarity of the constituent β-1,4-linked hexose backbones of these molecules. Thus, various fluorescent brightening agents and lectins bind to both cellulose and chitin. The identification of Acanthamoeba spp., which is based primarily on morphological and biochemical features, is labor-intensive and requires cloning and axenization. We describe a novel immunocytochemical method for identification of Acanthamoeba spp. based on selective binding of Trichoderma reesei cellulase to protozoan cyst wall cellulose. A recombinant cellulose-binding protein consisting of two cellulose-binding domains (CBDs) from T. reesei cellulases was coupled to the fluorescent dyes Alexa Fluor 350 and Alexa Fluor 568 or was labeled with biotin using EZ-Link sulfo-NHS-biotin. No staining reaction was observed with chitin-containing preparations of fungi. Thus, the recombinant CBDs can be used as a marker to distinguish between cellulose and chitin. This allows rapid identification of Acanthamoeba cyst wall cellulose in paraffin or frozen sections of infected tissues.

* Corresponding author. Mailing address: Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, 10 Fredry Street, 61-701 Poznan, Poland. Phone: 48 618546237. Fax: 48 618546236. E-mail: mderda{at}ump.edu.pl

{triangledown} Published ahead of print on 4 September 2009.

Applied and Environmental Microbiology, November 2009, p. 6827-6830, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.01555-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»