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Chem Biol. 2009 Jan 30;16(1):36-47. doi: 10.1016/j.chembiol.2008.11.004.

Novel fluorescent glycan microarray strategy reveals ligands for galectins.

Author information

1
Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA.

Abstract

Galectin-1 (Gal-1) and galectin-3 (Gal-3) are widely expressed galectins with immunoregulatory functions in animals. To explore their glycan specificity, we developed microarrays of naturally occurring glycans using a bifunctional fluorescent linker, 2-amino-N-(2-aminoethyl)-benzamide (AEAB), directly conjugated through its arylamine group by reductive amination to free glycans to form glycan-AEABs (GAEABs). Glycans from natural sources were used to prepare over 200 GAEABs, which were purified by multidimensional high-pressure liquid chromatography and covalently immobilized onto N-hydroxysuccinimide-activated glass slides via their free alkylamine. Fluorescence-based screening demonstrated that Gal-1 recognizes a wide variety of complex N-glycans, whereas Gal-3 primarily recognizes poly-N-acetyllactosamine-containing glycans independent of N-glycan presentation. GAEABs provide a general solution to glycan microarray preparation from natural sources for defining the specificity of glycan-binding proteins.

PMID:
19171304
PMCID:
PMC2662446
DOI:
10.1016/j.chembiol.2008.11.004
[Indexed for MEDLINE]
Free PMC Article

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