5KXC

Wisteria floribunda lectin in complex with GalNAc(beta1-4)GlcNAc (LacdiNAc) at pH 8.5.

Summary for 5KXC

• Entry DOI: 10.2210/pdb5kxc/pdb
• Related: 5KXB 5KXD 5KXE
• Descriptor: Wisteria floribunda agglutinin, alpha-L-fucopyranose-(1-3)-2-acetamido-2-deoxy-beta-D-glucopyranose, beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-3)]2-acetamido-2-deoxy-beta-D-glucopyranose, ... (8 entities in total)
• Functional Keywords: carbohydrate-binding protein, lectin, sugar binding protein
• Biological source: Wisteria floribunda
• Total number of polymer chains: 4
• Total formula weight: 110882.24

Authors

Evans, S.V.,Haji-Ghassemi, O. (deposition date: 2016-07-20, release date: 2016-09-14, Last modification date: 2024-11-06)

Primary citation

Haji-Ghassemi, O.,Gilbert, M.,Spence, J.,Schur, M.J.,Parker, M.J.,Jenkins, M.L.,Burke, J.E.,van Faassen, H.,Young, N.M.,Evans, S.V.
Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[ beta 14]GlcNAc), by Wisteria floribunda Agglutinin.
J.Biol.Chem., 291:24085-24095, 2016

PubMed Abstract: Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. The mechanism of specificity for WFA recognition of LacdiNAc is not fully understood. To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding K values of 4.67 × 10 m, 9.24 × 10 m, and 5.45 × 10 m, respectively. Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. Remarkably, the contribution of this small hydrophobic surface significantly increases the observed affinity for LacdiNAc over GalNAc. Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen).

PubMed: 27601469
DOI: 10.1074/jbc.M116.750463

Experimental method

X-RAY DIFFRACTION (1.8 Å)