Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis

Macarena Rodriguez-Walker, Aldo Alejandro Vilcaes, Eduardo Garbarino-Pico, José L. Daniotti

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Gangliosides are sialic acid-containing glycosphingolipids mainly expressed at the outer leaflet of the plasma membrane. Sialidase NEU3 is a key enzyme in the catabolism of gangliosides with its up-regulation having been observed in human cancer cells. In the case of CME (clathrin-mediated endocytosis), although this has been widely studied, the role of NEU3 and gangliosides in this cellular process has not yet been established. In the present study, we found an increased internalization of Tf (transferrin), the archetypical cargo for CME, in cells expressing complex gangliosides with high levels of sialylation. The ectopic expression of NEU3 led to a drastic decrease in Tf endocytosis, suggesting the participation of gangliosides in this process. However, the reduction in Tf endocytosis caused by NEU3 was still observed in glycosphingolipid-depleted cells, indicating that NEU3 could operate in a way that is independent of its action on gangliosides. Additionally, internalization of α2-macroglobulin and low-density lipoprotein, other typical ligands in CME, was also decreased in NEU3-expressing cells. In contrast, internalization of cholera toxin β-subunit, which is endocytosed by both clathrin-dependent and clathrin-independent mechanisms, remained unaltered. Kinetic assays revealed that NEU3 caused a reduction in the sorting of endocytosed Tf to early and recycling endosomes, with the Tf binding at the cell surface being also reduced. NEU3-expressing cells showed an altered subcellular distribution of clathrin adaptor AP-2 (adaptor protein 2), but did not reveal any changes in the membrane distribution of clathrin, PtdIns(4,5)P2 or caveolin-1. Overall, these results suggest a specific and novel role of NEU3 in CME.

Original languageEnglish (US)
Pages (from-to)131-144
Number of pages14
JournalBiochemical Journal
Volume470
Issue number1
DOIs
StatePublished - Aug 15 2015
Externally publishedYes

Fingerprint

Clathrin
Neuraminidase
Cell membranes
Endocytosis
Gangliosides
Transferrin
Cell Membrane
Glycosphingolipids
Vesicular Transport Adaptor Proteins
Phosphatidylinositol 4,5-Diphosphate
Cells
Caveolin 1
Macroglobulins
Cholera Toxin
N-Acetylneuraminic Acid
Endosomes
LDL Lipoproteins
Sorting
Recycling
Assays

Keywords

  • Clathrin
  • Ganglioside
  • Glycolipid
  • Membrane trafficking
  • NEU3
  • Sialidase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis. / Rodriguez-Walker, Macarena; Vilcaes, Aldo Alejandro; Garbarino-Pico, Eduardo; Daniotti, José L.

In: Biochemical Journal, Vol. 470, No. 1, 15.08.2015, p. 131-144.

Research output: Contribution to journalArticle

Rodriguez-Walker, M, Vilcaes, AA, Garbarino-Pico, E & Daniotti, JL 2015, 'Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis', Biochemical Journal, vol. 470, no. 1, pp. 131-144. https://doi.org/10.1042/BJ20141550
Rodriguez-Walker, Macarena ; Vilcaes, Aldo Alejandro ; Garbarino-Pico, Eduardo ; Daniotti, José L. / Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis. In: Biochemical Journal. 2015 ; Vol. 470, No. 1. pp. 131-144.
@article{4f7721317c3d41a5ae1d048a59ad2360,
title = "Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis",
abstract = "Gangliosides are sialic acid-containing glycosphingolipids mainly expressed at the outer leaflet of the plasma membrane. Sialidase NEU3 is a key enzyme in the catabolism of gangliosides with its up-regulation having been observed in human cancer cells. In the case of CME (clathrin-mediated endocytosis), although this has been widely studied, the role of NEU3 and gangliosides in this cellular process has not yet been established. In the present study, we found an increased internalization of Tf (transferrin), the archetypical cargo for CME, in cells expressing complex gangliosides with high levels of sialylation. The ectopic expression of NEU3 led to a drastic decrease in Tf endocytosis, suggesting the participation of gangliosides in this process. However, the reduction in Tf endocytosis caused by NEU3 was still observed in glycosphingolipid-depleted cells, indicating that NEU3 could operate in a way that is independent of its action on gangliosides. Additionally, internalization of α2-macroglobulin and low-density lipoprotein, other typical ligands in CME, was also decreased in NEU3-expressing cells. In contrast, internalization of cholera toxin β-subunit, which is endocytosed by both clathrin-dependent and clathrin-independent mechanisms, remained unaltered. Kinetic assays revealed that NEU3 caused a reduction in the sorting of endocytosed Tf to early and recycling endosomes, with the Tf binding at the cell surface being also reduced. NEU3-expressing cells showed an altered subcellular distribution of clathrin adaptor AP-2 (adaptor protein 2), but did not reveal any changes in the membrane distribution of clathrin, PtdIns(4,5)P2 or caveolin-1. Overall, these results suggest a specific and novel role of NEU3 in CME.",
keywords = "Clathrin, Ganglioside, Glycolipid, Membrane trafficking, NEU3, Sialidase",
author = "Macarena Rodriguez-Walker and Vilcaes, {Aldo Alejandro} and Eduardo Garbarino-Pico and Daniotti, {Jos{\'e} L.}",
year = "2015",
month = "8",
day = "15",
doi = "10.1042/BJ20141550",
language = "English (US)",
volume = "470",
pages = "131--144",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "1",

}

TY - JOUR

T1 - Role of plasma-membrane-bound sialidase NEU3 in clathrin-mediated endocytosis

AU - Rodriguez-Walker, Macarena

AU - Vilcaes, Aldo Alejandro

AU - Garbarino-Pico, Eduardo

AU - Daniotti, José L.

PY - 2015/8/15

Y1 - 2015/8/15

N2 - Gangliosides are sialic acid-containing glycosphingolipids mainly expressed at the outer leaflet of the plasma membrane. Sialidase NEU3 is a key enzyme in the catabolism of gangliosides with its up-regulation having been observed in human cancer cells. In the case of CME (clathrin-mediated endocytosis), although this has been widely studied, the role of NEU3 and gangliosides in this cellular process has not yet been established. In the present study, we found an increased internalization of Tf (transferrin), the archetypical cargo for CME, in cells expressing complex gangliosides with high levels of sialylation. The ectopic expression of NEU3 led to a drastic decrease in Tf endocytosis, suggesting the participation of gangliosides in this process. However, the reduction in Tf endocytosis caused by NEU3 was still observed in glycosphingolipid-depleted cells, indicating that NEU3 could operate in a way that is independent of its action on gangliosides. Additionally, internalization of α2-macroglobulin and low-density lipoprotein, other typical ligands in CME, was also decreased in NEU3-expressing cells. In contrast, internalization of cholera toxin β-subunit, which is endocytosed by both clathrin-dependent and clathrin-independent mechanisms, remained unaltered. Kinetic assays revealed that NEU3 caused a reduction in the sorting of endocytosed Tf to early and recycling endosomes, with the Tf binding at the cell surface being also reduced. NEU3-expressing cells showed an altered subcellular distribution of clathrin adaptor AP-2 (adaptor protein 2), but did not reveal any changes in the membrane distribution of clathrin, PtdIns(4,5)P2 or caveolin-1. Overall, these results suggest a specific and novel role of NEU3 in CME.

AB - Gangliosides are sialic acid-containing glycosphingolipids mainly expressed at the outer leaflet of the plasma membrane. Sialidase NEU3 is a key enzyme in the catabolism of gangliosides with its up-regulation having been observed in human cancer cells. In the case of CME (clathrin-mediated endocytosis), although this has been widely studied, the role of NEU3 and gangliosides in this cellular process has not yet been established. In the present study, we found an increased internalization of Tf (transferrin), the archetypical cargo for CME, in cells expressing complex gangliosides with high levels of sialylation. The ectopic expression of NEU3 led to a drastic decrease in Tf endocytosis, suggesting the participation of gangliosides in this process. However, the reduction in Tf endocytosis caused by NEU3 was still observed in glycosphingolipid-depleted cells, indicating that NEU3 could operate in a way that is independent of its action on gangliosides. Additionally, internalization of α2-macroglobulin and low-density lipoprotein, other typical ligands in CME, was also decreased in NEU3-expressing cells. In contrast, internalization of cholera toxin β-subunit, which is endocytosed by both clathrin-dependent and clathrin-independent mechanisms, remained unaltered. Kinetic assays revealed that NEU3 caused a reduction in the sorting of endocytosed Tf to early and recycling endosomes, with the Tf binding at the cell surface being also reduced. NEU3-expressing cells showed an altered subcellular distribution of clathrin adaptor AP-2 (adaptor protein 2), but did not reveal any changes in the membrane distribution of clathrin, PtdIns(4,5)P2 or caveolin-1. Overall, these results suggest a specific and novel role of NEU3 in CME.

KW - Clathrin

KW - Ganglioside

KW - Glycolipid

KW - Membrane trafficking

KW - NEU3

KW - Sialidase

UR - http://www.scopus.com/inward/record.url?scp=84938801354&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938801354&partnerID=8YFLogxK

U2 - 10.1042/BJ20141550

DO - 10.1042/BJ20141550

M3 - Article

C2 - 26251452

AN - SCOPUS:84938801354

VL - 470

SP - 131

EP - 144

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 1

ER -