Glycan array analysis of Pholiota squarrosa lectin (PhoSL) and other fucose-oriented lectins.

Glycan array analysis of Pholiota squarrosa lectin (PhoSL) and other fucose-oriented lectins.

Glycobiology. 2020 Oct 06;:

Authors: Rubén LC, Laura MR, Almudena FB, Emilio GM

The α(1,6)fucose residue attached to the N-glycoprotein core is suspected to play an essential role in the progression of several types of cancer. Lectins remain the first choice for probing glycan modifications, although they may lack specificity. Thus, efforts have been made to identify new lectins with a narrower core fucose detection profile. Here, we present a comparison of the classical Aleuria aurantia lectin (AAL), Lens culinaris agglutinin (LCA) and Aspergillus oryzae lectin (AOL) with the newer Pholiota squarrosa lectin (PhoSL), which has been described as being specific for core fucosylated N-glycans. To this end, we studied the binding profiles of the four lectins using mammalian glycan arrays from the Consortium of Functional Glycomics. To validate their glycan specificity, we probed AOL, LCA and PhoSL in western-blot assays using protein extracts from eight common colorectal cancer lines and colorectal biopsies from a small cohort of patients with colorectal cancer. The results showed that i) LCA and PhoSL were the most specific lectins for detecting the presence of core fucose in a concentration-dependent manner; ii) PhoSL exhibited the highest N-glycan sequence restriction, with preferential binding to core fucosylated paucimannosidic-type N-glycans, iii) the recognition ability of PhoSL was highly influenced by the presence of terminal N-acetyl-lactosamine; iv) LCA bound to paucimannosidic, bi-antennary and tri-antennary core fucosylated N-glycans; and v) AOL and AAL exhibited broader specificity towards fucosylation. Together, our results support the choice of LCA as the most appropriate lectin for core fucose detection, as validated in protein extracts from colorectal cancer cell lines and tissue specimens from patients with colorectal cancer.

PMID: 33021632 [PubMed – as supplied by publisher]

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