Sialidase NEU3 is a peripheral membrane protein localized on the cell surface and in endosomal structures.
Sialidase NEU3 is also known as the plasma-membrane-associated form of mammalian sialidases, exhibiting a high substrate specificity towards gangliosides. In this respect, sialidase NEU3 modulates cell-surface biological events and plays a pivotal role in different cellular processes, including cell adhesion, recognition and differentiation. At the moment, no detailed studies concerning the subcellular localization of NEU3 are available, and the mechanism of its association with cellular membranes is still unknown. In the present study, we have demonstrated that sialidase NEU3, besides its localization at the plasma membrane, is present in intracellular structures at least partially represented by a subset of the endosomal compartment. Moreover, we have shown that NEU3 present at the plasma membrane is internalized and locates then to the recycling endosomal compartment. The enzyme is associated with the outer leaflet of the plasma membrane, as shown by selective cell-surface protein biotinylation. This evidence is in agreement with the ability of NEU3 to degrade gangliosides inserted into the plasma membrane of adjacent cells. Moreover, the mechanism of the protein association with the lipid bilayer was elucidated by carbonate extraction. Under these experimental conditions, we have succeeded in solubilizing NEU3, thus demonstrating that the enzyme is a peripheral membrane protein. In addition, Triton X-114 phase separation demonstrates further the hydrophilic nature of the protein. Overall, these results provide important information about the biology of NEU3, the most studied member of the mammalian sialidase family.
Zanchetti
Gabriele
G
Department of Biomedical Sciences and Biotechnology, School of Medicine, University of Brescia, Viale Europa 11, 25123 Brescia, Italy.
Colombi
Paolo
P
Manzoni
Marta
M
Anastasia
Luigi
L
Caimi
Luigi
L
Borsani
Giuseppe
G
Venerando
Bruno
B
Tettamanti
Guido
G
Preti
Augusto
A
Monti
Eugenio
E
Bresciani
Roberto
R
D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000201:enzymology* / Q000378:metabolism; D002522:Chlorocebus aethiops; D011992:Endosomes / Q000201:enzymology* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism*
D008565:Membrane Proteins; C465436:Neu3 protein, human; D009439:Neuraminidase
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