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Elucidating the selectivity of recombinant forms of Aleuria aurantia lectin using weak affinity chromatography
Linnaeus University.
Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
Linnaeus University.
2012 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 885, 66-72 p.Article in journal (Refereed) Published
Abstract [en]

Aberrant glycosylation is connected to several pathological conditions and lectins are useful tools to characterize glycosylated biomarkers. The Aleuria aurantia lectin (AAL) is of special interest since it interacts with all types of fucosylated saccharides. AAL has been expressed in Escherichia coil as a fully functional recombinant protein. Engineered variants of AAL have been developed with the aim of creating monovalent lectins with more homogenous binding characteristics. Four different forms of AAL were studied in the present work: native AAL purified from A. aurantia mushrooms, recombinant AAL dimer, recombinant AAL monomer and recombinant AAL site 2 (S2-AAL). The affinities of these AAL forms toward a number of saccharides were determined with weak affinity chromatography (WAC). Disaccharides with fucose linked alpha 1-3 to GIcNAc interacted with higher affinity compared to fucose linked alpha 1-6 or alpha 1-4 and the obtained dissociation constants (K-d) were in the range of 10 mu M for all AAL forms. Tetra- and pentasaccharides with fucose in alpha 1-2, alpha 1-3 or alpha 1-4 had K-d values ranging from 0.1 to 7 mM while a large alpha 1-6 fucosylated oligosaccharide had a K-d of about 20 mu M. The recombinant multivalent AAL forms and native AAL exhibited similar affinities toward all saccharides, but S2-AAL had a lower affinity especially regarding a sialic acid containing fucosylated saccharide. It was demonstrated that WAC is a valuable technique in determining the detailed binding profile of the lectins. Specific advantages with WAC include a low consumption of non-labeled saccharides, possibility to analyze mixtures and a simple procedure using standard HPLC equipment.

Place, publisher, year, edition, pages
Elsevier , 2012. Vol. 885, 66-72 p.
Keyword [en]
Affinity, Aleuria aurantia lectin, Glycan interaction, Recombinant protein, Weak affinity chromatography
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-76625DOI: 10.1016/j.jchromb.2011.12.015ISI: 000301563400010OAI: diva2:515430
Funding Agencies|Linnaeus University||Linkoping University||Medical council of Southeast Sweden||Available from: 2012-04-13 Created: 2012-04-13 Last updated: 2012-06-05

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Åström, EvaPåhlsson, Peter
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