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. 2014 Aug 29;19(3):461–482. doi: 10.2478/s11658-014-0206-4

The lectin-binding pattern of nucleolin and its interaction with endogenous galectin-3

Dorota Hoja-Łukowicz 1,✉, Sylwia Kedracka-Krok 2,3, Weronika Duda 4, Anna Lityńska 1
PMCID: PMC6275868  PMID: 25169435

Abstract

Unlike nuclear nucleolin, surface-expressed and cytoplasmic nucleolin exhibit Tn antigen. Here, we show localization-dependent differences in the glycosylation and proteolysis patterns of nucleolin. Our results provide evidence for different paths of nucleolin proteolysis in the nucleus, in the cytoplasm, and on the cell surface. We found that full-length nucleolin and some proteolytic fragments coexist within live cells and are not solely the result of the preparation procedure. Extranuclear nucleolin undergoes N- and O-glycosylation, and unlike cytoplasmic nucleolin, membrane-associated nucleolin is not fucosylated. Here, we show for the first time that nucleolin and endogenous galectin-3 exist in the same complexes in the nucleolus, the cytoplasm, and on the cell surface of melanoma cells. Assessments of the interaction of nucleolin with galectin-3 revealed nucleolar co-localization in interphase, suggesting that galectin-3 may be involved in DNA organization and ribosome biogenesis.

Electronic Supplementary Material

Supplementary material is available for this article at 10.2478/s11658-014-0206-4 and is accessible for authorized users.

Keywords: Glycosylation of nucleolin, Galectin-3, Melanoma, Mass spectrometry, Confocal microscopy, Lectin assay, Co-immunoprecipitation

Full Text

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Abbreviations used

AAA

Aleuria aurantia agglutinin

DSA

Datura stramonium gglutinin

GNA

Galanthus nivalis agglutinin

H antigen

Fucα1-2Galβ1-4GlcNAcβ1-3Gal-

LEA

Lycopersicon esculentum agglutinin

Lex

Galβ1-4(Fucα1-3)GlcNAcβ-

LTA

Tetragonolobus purpureus agglutinin

MAA-II

Maackia amurensis agglutinin

SNA-I

Sambucus nigra agglutinin

PHA-E

Phaseolus vulgaris erythroagglutinin

PHAL

Phaseolus vulgaris leucoagglutinin

PNA

peanut agglutinin

T antigen

Galβ1-3GalNAcα1 -Ser/Thr

Tn antigen

GalNAcα1 -Ser/Thr

UEA-I

Ulex europaeus agglutinin

VVA

Vicia villosa agglutinin

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