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

Communications

Fishing for Lectins from Diverse Sequence Libraries by Yeast Surface Display – An Exploratory Study

Stefan Ryckaert1,2,*, Nico Callewaert3,4,*, Pieter P. Jacobs1,2,, Sylviane Dewaele1,2,, Isabelle Dewerte3,4, and Roland Contreras1,2,

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

Corresponding author: Nico Callewaert, Unit for Molecular Glycobiology, FSVM research building, Technologiepark 927, B-9052 Gent-Zwijnaarde, Belgium. Tel. +32 9 331 3617, Fax. +32 9 331 3502, E-mail: Nico.Callewaert{at}DMBR.UGent.be

Received on May 9, 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, fluorescently labelled polyacrylamide-based glycoconjugates.

Key words: Expression Cloning / Lectin / Pichia pastoris / Saccharomyces cerevisiae / Yeast Surface Display

* Both first authors contributed equally.


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