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

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.

Abbreviations

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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Hoja-Łukowicz, D., Kedracka-Krok, S., Duda, W. et al. The lectin-binding pattern of nucleolin and its interaction with endogenous galectin-3. Cell Mol Biol Lett 19, 461–482 (2014). https://doi.org/10.2478/s11658-014-0206-4

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  • DOI: https://doi.org/10.2478/s11658-014-0206-4

Keywords

  • Glycosylation of nucleolin
  • Galectin-3
  • Melanoma
  • Mass spectrometry
  • Confocal microscopy
  • Lectin assay
  • Co-immunoprecipitation