Terminal N-acetylgalactosamine-specific leguminous lectin from Wisteria japonica as a probe for human lung squamous cell carcinoma

PloS One
Keisuke SogaKazuo Yamamoto

Abstract

Millettia japonica was recently reclassified into the genus Wisteria japonica based on chloroplast and nuclear DNA sequences. Because the seed of Wisteria floribunda expresses leguminous lectins with unique N-acetylgalactosamine-binding specificity, we purified lectin from Wisteria japonica seeds using ion exchange and gel filtration chromatography. Glycan microarray analysis demonstrated that unlike Wisteria floribunda and Wisteria brachybotrys lectins, which bind to both terminal N-acetylgalactosamine and galactose residues, Wisteria japonica lectin (WJA) specifically bound to both α- and β-linked terminal N-acetylgalactosamine, but not galactose residues on oligosaccharides and glycoproteins. Further, frontal affinity chromatography using more than 100 2-aminopyridine-labeled and p-nitrophenyl-derivatized oligosaccharides demonstrated that the ligands with the highest affinity for Wisteria japonica lectin were GalNAcβ1-3GlcNAc and GalNAcβ1-4GlcNAc, with K(a) values of 9.5 × 10(4) and 1.4 × 10(5) M(-1), respectively. In addition, when binding was assessed in a variety of cell lines, Wisteria japonica lectin bound specifically to EBC-1 and HEK293 cells while other Wisteria lectins bound equally to all of the cell lines tested....Continue Reading

References

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Citations

May 31, 2017·International Journal of Molecular Sciences·Yoko ItakuraJun Hirabayashi
Jan 13, 2015·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jun HirabayashiHisashi Narimatsu
Jan 9, 2020·Biochimica Et Biophysica Acta. General Subjects·Martina PirroPaul J Hensbergen
Aug 27, 2018·International Journal of Biological Macromolecules·Ana Paula M NascimentoRodrigo B Leal

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Methods Mentioned

BETA
glycosylation
ion
gel filtration
electrophoresis
flow cytometry
FACS
PCR

Software Mentioned

CellQuest
FAC

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