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Glycobiology Advance Access originally published online on December 17, 2007
Glycobiology 2008 18(2):137-144; doi:10.1093/glycob/cwm131
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© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Communication

Fishing for lectins from diverse sequence libraries by yeast surface display – An exploratory study

Stefan Ryckaert2,3,4,, Nico Callewaert1,5,6,, Pieter P. Jacobs3,4,, Sylviane Dewaele3,4,, Isabelle Dewerte5,6, and Roland Contreras3,4,

3 Department for Molecular Biomedical Research, Unit for Fundamental and Applied Molecular Biology, VIB
4 Department of Molecular Biology, Ghent University
5 Department for Molecular Biomedical Research, Unit for Molecular Glycobiology, VIB
6 Department of Biochemistry, Physiology and Microbiology, Ghent University, B-9052 Ghent, Belgium

1 To whom correspondence should be addressed: Tel: +32-9-331-3617; Fax: +32-9-331-3502; e-mail: Nico.Callewaert{at}DMBR.UGent.be

Received on May 9, 2007; revised on November 28, 2007; accepted on November 30, 2007

The establishment of a robust technology platform for the expression cloning of carbohydrate-binding proteins remains a key challenge in glycomics. Here we explore the utility of using yeast surface display (YSD) technology in the interaction-based lectin cloning from complete cDNA libraries. This should pave the way for more detailed studies of protein–carbohydrate interactions. To evaluate the performance of this system, lectins representing three different subfamilies (galectins, siglecs, and C-type lectins) were successfully displayed on the surface of Saccharomyces cerevisiae and Pichia pastoris as a-agglutinin and/or {alpha}-agglutinin fusions. The predicted carbohydrate-binding activity could be detected for three out of five lectins tested (galectin-1, galectin-3, and siaoadhesin). For galectin-4 and E-selectin, no specific carbohydrate-binding activity could be detected. We also demonstrate that proteins with carbohydrate affinity can be specifically isolated from complex metazoan cDNA libraries through multiple rounds of FACS sorting, employing multivalent, fluorescent-labeled polyacrylamide-based glycoconjugates.

Key words: expression cloning / lectin / Pichia pastoris / Saccharomyces cerevisiae / yeast surface display

2 Both first authors contributed equally.


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