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Exquisite binding specificity of Sclerotium rolfsii lectin toward TF-related O-linked mucin-type glycans

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Abstract

Sclerotium rolfsii lectin (SRL), a secretory protein from the soil borne phytopathogenic fungus Sclerotium rolfsii, has shown in our previous studies to bind strongly to the oncofetal Thomson-Friedenreich carbohydrate (Galβ1-3GalNAc-ser/thr, T or TF) antigen. TF antigen is widely expressed in many types of human cancers and the strong binding of SRL toward such a cancer-associated carbohydrate structure led us to characterize the carbohydrate binding specificity of SRL. Glycan array analysis, which included 285 glycans, shows exclusive binding of SRL to the O-linked mucin type but not N-linked glycans and amongst the mucin type O-glycans, lectin recognizes only mucin core 1, core 2 and weakly core 8 but not to other mucin core structures. It binds with high specificity to “α-anomers” but not the “β-anomers” of the TF structure. The axial C4-OH group of GalNAc and C2-OH group of Gal is both essential for SRL interaction with TF disaccharide, and substitution on C3 of galactose by sulfate or sialic acid or N-acetylglucosamine, significantly enhances the avidity of the lectin. SRL differs in its binding to TF structures compared to other known TF-binding lectins such as the Arachis hypogea (peanut) agglutinin, Agaricus bisporus (mushroom) lectin, Jackfruit, Artocarpus integrifolia (jacalin) and Amaranthus caudatus (Amaranthin) lectin. Thus, SRL has unique carbohydrate-binding specificity toward TF-related O-linked carbohydrate structures. Such a binding specificity will make this lectin a very useful tool in future structural as well as functional analysis of the cellular glycans in cancer studies.

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Abbreviations

ABL:

Agaricus bisporus lectin

ACA:

Amaranthus caudatus agglutinin

AGP:

α-1 Acid glycoprotein

ELLA:

Enzyme-Linked Lectinosorbent assay

GalNAc:

N-acetylgalactosamine

GBP:

Glycan Binding Proteins

GIPC:

Glycosyl Inositol Phosphoryl Ceramide

GlcNAc:

N-acetylglucosamine

G:

Glycan

GM1:

Ganglioside GM1

LacNAc:

N-acetyllactosamine

Neu5Ac:

N-acetylneuraminic acid

PNA:

Peanut agglutinin

RFU:

Relative Fluorescence Units

SRL:

Sclerotium rolfsii lectin

TF:

Thomson Friedenreich

core 1:

Galβ1-3GalNAcα

core 2:

Galβ1-3 (GlcNAcβ1-6) GalNAcα

core 3:

GlcNAcβ1-3GalNAcα

core 4:

GlcNAcβ1-3 (GlcNAcβ1-6) GalNAcα

core 6:

GlcNAcβ1-6GalNAcα

core 8:

Galα 1-3GalNAcα

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Acknowledgement

We would like to thank the Consortium for Functional Glycomics, for the glycan array analysis. This work was supported by the Department of Science and Technology (DST), New Delhi (Grant No-SR/SO/BB/43/2003) and partially by British council under UKIERI program.

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Authors and Affiliations

  1. P.G. Department of Studies in Biochemistry, Karnatak University, Dharwad, 580 003, India

    Vishwanath B. Chachadi, Shashikala R. Inamdar & Bale M. Swamy

  2. The Henry Wellcome Laboratory of Medicine and cellular gastroenterology, School of Clinical Sciences, University of Liverpool, Liverpool, L69 3BX, UK

    Lu-Gang Yu & Jonathan M. Rhodes

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  1. Vishwanath B. Chachadi
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  2. Shashikala R. Inamdar
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  3. Lu-Gang Yu
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  5. Bale M. Swamy
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Correspondence to Bale M. Swamy.

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Chachadi, V.B., Inamdar, S.R., Yu, LG. et al. Exquisite binding specificity of Sclerotium rolfsii lectin toward TF-related O-linked mucin-type glycans. Glycoconj J 28, 49–56 (2011). https://doi.org/10.1007/s10719-011-9323-8

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  • DOI: https://doi.org/10.1007/s10719-011-9323-8

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